Substituted piperazines as selective hdac1,2 inhibitors

ABSTRACT

Provided herein are compounds, pharmaceutical compositions comprising such compounds, and methods of using such compounds to treat diseases or disorders associated with HDAC1 and/or HDAC2 activity.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/425,938, filed Nov. 23, 2016, and U.S. Provisional Application No.62/506,344, filed May 15, 2017. The entire contents of theseapplications are incorporated herein in their entireties.

BACKGROUND

A biological target of current interest is histone deacetylase (HDAC)(see, for example, a discussion of the use of inhibitors of histonedeacetylases for the treatment of cancer: Marks et al. Nature ReviewsCancer 2001, 7, 194; Johnstone et al. Nature Reviews Drug Discovery2002, 287). Post-translational modification of proteins throughacetylation and deacetylation of lysine residues plays a critical rolein regulating their cellular functions. HDACs are zinc hydrolases thatmodulate gene expression through deacetylation of the N-acetyl-lysineresidues of histone proteins and other transcriptional regulators(Hassig et al. Curr. Opin. Chem. Biol. 1997, 1, 300-308). HDACsparticipate in cellular pathways that control cell shape anddifferentiation, and an HDAC inhibitor has been shown to be effective intreating an otherwise recalcitrant cancer (Warrell et al. J. Natl.Cancer Inst. 1998, 90, 1621-1625).

Eleven human HDACs, which use Zn as a cofactor, have been identified(Taunton et al. Science 1996, 272, 408-411; Yang et al. J. Biol. Chem.1997, 272, 28001-28007. Grozinger et al. Proc. Natl. Acad. Sci. U.S.A.1999, 96, 4868-4873; Kao et al. Genes Dev. 2000, 14, 55-66. Hu et al. J.Biol. Chem. 2000, 275, 15254-15264; Zhou et al. Proc. Natl. Acad. SciU.S.A. 2001, 98, 10572-10577; Venter et al. Science 2001, 291,1304-1351) and these members fall into three classes (class I, II, andIV) based on sequence homology to their yeast orthologues (O. Witt etal. Cancer Letters, 2009, 277, 8-21). Class I HDACs include HDAC1,HDAC2, HDAC3, and HDAC8, and are referred to as “classical” HDACs, whichimplies a catalytic pocket with a Zn²⁺ ion at its base.

There remains a need for preparing structurally diverse HDAC inhibitors,particularly ones that are potent and/or selective inhibitors ofparticular classes of HDACs and individual HDACs.

SUMMARY

Provided herein are compounds and methods of using these compounds totreat disorders related to HDAC1 or HDAC2 function, including cancer,myelodysplastic syndrome or hemoglobinopathy.

Thus, in an aspect, provided herein are compounds of Formula I:

or a pharmaceutically acceptable salt thereof.

Also provided herein is a pharmaceutical composition comprising acompound of Formula I, or a pharmaceutically acceptable salt thereof,together with a pharmaceutically acceptable carrier.

In another aspect, provided herein are compounds of Formula II:

or a pharmaceutically acceptable salt thereof.

Also provided herein is a pharmaceutical composition comprising acompound of Formula II, or a pharmaceutically acceptable salt thereof,together with a pharmaceutically acceptable carrier.

In a further aspect, provided here are compounds of Formula III:

or a pharmaceutically acceptable salt thereof.

Also provided herein is a pharmaceutical composition comprising acompound of Formula III, or a pharmaceutically acceptable salt thereof,together with a pharmaceutically acceptable carrier.

In yet another aspect, provided herein are compounds of Formula IV:

or a pharmaceutically acceptable salt thereof.

Also provided herein is a pharmaceutical composition comprising acompound of Formula III, or a pharmaceutically acceptable salt thereof,together with a pharmaceutically acceptable carrier.

In an aspect, provided herein is a method of inhibiting the activity ofHDAC1 and/or HDAC2 in a subject in need thereof, comprisingadministering to the subject a compound of Formula I, or apharmaceutically acceptable salt thereof.

In an aspect, provided herein is a method of inhibiting the activity ofHDAC1 and/or HDAC2 in a subject in need thereof, comprisingadministering to the subject a compound of Formula II, Formula III,Formula IV, or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method of treating a diseasemediated by HDAC1 and/or HDAC2 in a subject in need thereof, comprisingadministering to the subject a compound of Formula I, or apharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method of treating a diseasemediated by HDAC1 and/or HDAC2 in a subject in need thereof, comprisingadministering to the subject a compound of Formula II, Formula III,Formula IV, or a pharmaceutically acceptable salt thereof.

In yet another aspect, provided herein is a method of treating amyelodysplastic syndrome in a subject in need thereof, comprisingadministering to the subject a compound of Formula I, or apharmaceutically acceptable salt thereof.

In yet another aspect, provided herein is a method of treating amyelodysplastic syndrome in a subject in need thereof, comprisingadministering to the subject a compound of Formula II, Formula III,Formula IV, or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method of treating ahemoglobinopathy in a subject in need thereof, comprising administeringto the subject a compound of Formula I, or a pharmaceutically acceptablesalt thereof. In an embodiment, the hemoglobinopathy is sickle-celldisease or beta-thalassemia.

In another aspect, provided herein is a method of treating ahemoglobinopathy in a subject in need thereof, comprising administeringto the subject a compound of Formula II, Formula III, Formula IV, or apharmaceutically acceptable salt thereof. In an embodiment, thehemoglobinopathy is sickle-cell disease or beta-thalassemia.

In another aspect, provided herein is a method of treating a cancer in asubject in need thereof, comprising administering to the subject acompound of Formula I, or a pharmaceutically acceptable salt thereof. Inan embodiment, the cancer is lung cancer, colon cancer, breast cancer,neuroblastoma, leukemia, or lymphoma. In a further embodiment, thecancer is acute myelogenous leukemia or acute megakaryocytic leukemia.In yet another embodiment, the cancer is neuroblastoma.

In another aspect, provided herein is a method of treating a cancer in asubject in need thereof, comprising administering to the subject acompound of Formula II, Formula III, Formula IV, or a pharmaceuticallyacceptable salt thereof. In an embodiment, the cancer is lung cancer,colon cancer, breast cancer, neuroblastoma, leukemia, or lymphoma. In afurther embodiment, the cancer is acute myelogenous leukemia or acutemegakaryocytic leukemia. In yet another embodiment, the cancer isneuroblastoma.

In a further aspect, provided herein is a method for treating sicklecell disease, beta thalassemia, myelodysplastic syndrome, acutemyelogenous leukemia, neuroblastoma, or acute megakaryocytic leukemia ina subject in need thereof, comprising administering to the subject atherapeutically effective amount of a compound of a compound of FormulaI, or a pharmaceutically acceptable salt thereof.

In a further aspect, provided herein is a method for treating sicklecell disease, beta thalassemia, myelodysplastic syndrome, acutemyelogenous leukemia, neuroblastoma, or acute megakaryocytic leukemia ina subject in need thereof, comprising administering to the subject atherapeutically effective amount of a compound of a compound of FormulaII, Formula III, Formula IV, or a pharmaceutically acceptable saltthereof.

DETAILED DESCRIPTION

Provided herein are compounds, e.g., the compounds of Formula I orpharmaceutically acceptable salts thereof, that are useful in thetreatment of cancer, myelodysplastic syndrome or hemoglobinopathy in asubject.

In a non-limiting aspect, these compounds can inhibit histonedeacetylases. In a particular embodiment, the compounds provided hereinare considered HDAC1 and/or HDAC2 inhibitors. As such, in one aspect,the compounds provided herein are useful in the treatment of cancer,myelodysplastic syndrome or hemoglobinopathy in a subject by acting as aHDAC1 and/or HDAC2 inhibitor.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

Unless defined otherwise, all technical and scientific terms used hereingenerally have the same meaning as commonly understood by one ofordinary skill in the art to which this invention belongs. Generally,the nomenclature used herein and the laboratory procedures in cellculture, molecular genetics, organic chemistry, and peptide chemistryare those well-known and commonly employed in the art.

As used herein, the articles “a” and “an” refer to one or to more thanone (i.e., to at least one) of the grammatical object of the article. Byway of example, “an element” means one element or more than one element.Furthermore, use of the term “including” as well as other forms, such as“include,” “includes,” and “included,” is not limiting.

As used herein, the term “about” will be understood by persons ofordinary skill in the art and will vary to some extent on the context inwhich it is used. As used herein when referring to a measurable valuesuch as an amount, a temporal duration, and the like, the term “about”is meant to encompass variations of ±20% or ±10%, including ±5%, ±1%,and ±0.1% from the specified value, as such variations are appropriateto perform the disclosed methods.

The term “treat,” “treated,” “treating,” or “treatment” includes thediminishment or alleviation of at least one symptom associated or causedby the state, disorder or disease being treated. In certain embodiments,the treatment comprises bringing into contact with HDAC1 and/or HDAC2 aneffective amount of a compound of the invention for conditions relatedto cancers, hemoglobinopathies, or myelodysplastic syndrome.

As used herein, the term “prevent” or “prevention” means no disorder ordisease development if none had occurred, or no further disorder ordisease development if there had already been development of thedisorder or disease. Also considered is the ability of one to preventsome or all of the symptoms associated with the disorder or disease.

The phrase “jointly therapeutically active” or “joint therapeuticeffect” as used herein means that the therapeutic agents can be givenseparately (in a chronologically staggered manner, especially asequence-specific manner) in such time intervals that they prefer, inthe warm-blooded animal, especially human, to be treated, still show an(preferably synergistic) interaction (joint therapeutic effect). Whetherthis is the case can, inter alia, be determined by following the bloodlevels of the compounds, showing that both compounds are present in theblood of the human to be treated at least during certain time intervals.

As used herein, the term “resistant” or “refractive” to a therapeuticagent when referring to a cancer patient means that the cancer hasinnate, or achieved resistance to, the effects of the therapeutic agentas a result of contact with the therapeutic agent. Stated alternatively,the cancer is resistant to the ordinary standard of care associated withthe particular therapeutic agent.

As used herein, the term “patient,” “individual,” or “subject” refers toa human or a non-human mammal. Non-human mammals include, for example,livestock and pets, such as ovine, bovine, porcine, canine, feline andmarine mammals. Preferably, the patient, subject, or individual ishuman.

As used herein, the terms “effective amount,” “pharmaceuticallyeffective amount,” and “therapeutically effective amount” refer to anontoxic but sufficient amount of an agent to provide the desiredbiological result. That result may be reduction or alleviation of thesigns, symptoms, or causes of a disease, or any other desired alterationof a biological system. An appropriate therapeutic amount in anyindividual case may be determined by one of ordinary skill in the artusing routine experimentation.

As used herein, the term “pharmaceutically acceptable” refers to amaterial, such as a carrier or diluent, which does not abrogate thebiological activity or properties of the compound, and is relativelynon-toxic, i.e., the material may be administered to an individualwithout causing undesirable biological effects or interacting in adeleterious manner with any of the components of the composition inwhich it is contained.

As used herein, the term “pharmaceutically acceptable salt” refers toderivatives of the disclosed compounds wherein the parent compound ismodified by converting an existing acid or base moiety to its salt form.Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts of thepresent invention include the conventional non-toxic salts of the parentcompound formed, for example, from non-toxic inorganic or organic acids.The pharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, nonaqueousmedia like ether, ethyl acetate, ethanol, isopropanol, or acetonitrileare preferred. The phrase “pharmaceutically acceptable salt” is notlimited to a mono, or 1:1, salt. For example, “pharmaceuticallyacceptable salt” also includes bis-salts, such as a bis-hydrochloridesalt. Lists of suitable salts are found in Remington's PharmaceuticalSciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418and Journal of Pharmaceutical Science, 66, 2 (1977), each of which isincorporated herein by reference in its entirety.

As used herein, the term “composition” or “pharmaceutical composition”refers to a mixture of at least one compound useful within the inventionwith a pharmaceutically acceptable carrier. The pharmaceuticalcomposition facilitates administration of the compound to a patient orsubject. Multiple techniques of administering a compound exist in theart including, but not limited to, intravenous, oral, aerosol,parenteral, ophthalmic, pulmonary, and topical administration.

As used herein, the term “pharmaceutically acceptable carrier” means apharmaceutically acceptable material, composition or carrier, such as aliquid or solid filler, stabilizer, dispersing agent, suspending agent,diluent, excipient, thickening agent, solvent or encapsulating material,involved in carrying or transporting a compound useful within theinvention within or to the patient such that it may perform its intendedfunction. Typically, such constructs are carried or transported from oneorgan, or portion of the body, to another organ, or portion of the body.Each carrier must be “acceptable” in the sense of being compatible withthe other ingredients of the formulation, including the compound usefulwithin the invention, and not injurious to the patient. Some examples ofmaterials that may serve as pharmaceutically acceptable carriersinclude: sugars, such as lactose, glucose and sucrose; starches, such ascorn starch and potato starch; cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients, such as cocoabutter and suppository waxes; oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols,such as propylene glycol; polyols, such as glycerin, sorbitol, mannitoland polyethylene glycol; esters, such as ethyl oleate and ethyl laurate;agar; buffering agents, such as magnesium hydroxide and aluminumhydroxide; surface active agents; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffersolutions; and other non-toxic compatible substances employed inpharmaceutical formulations.

As used herein, “pharmaceutically acceptable carrier” also includes anyand all coatings, antibacterial and antifungal agents, and absorptiondelaying agents, and the like that are compatible with the activity ofthe compound useful within the invention, and are physiologicallyacceptable to the patient. Supplementary active compounds may also beincorporated into the compositions. The “pharmaceutically acceptablecarrier” may further include a pharmaceutically acceptable salt of thecompound useful within the invention. Other additional ingredients thatmay be included in the pharmaceutical compositions used in the practiceof the invention are known in the art and described, for example, inRemington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co.,1985, Easton, Pa.), which is incorporated herein by reference.

The terms “combination,” “therapeutic combination,” or “pharmaceuticalcombination” as used herein refer to either a fixed combination in onedosage unit form, or non-fixed combination, or a kit of parts for thecombined administration where two or more therapeutic agents may beadministered independently, at the same time or separately within timeintervals, especially where these time intervals allow that thecombination partners show a cooperative, e.g., synergistic, effect.

The term “combination therapy” refers to the administration of two ormore therapeutic agents to treat a therapeutic condition or disorderdescribed in the present disclosure. Such administration encompassesco-administration of these therapeutic agents in a substantiallysimultaneous manner, such as in a single formulation having a fixedratio of active ingredients or in separate formulations (e.g., capsulesand/or intravenous formulations) for each active ingredient. Inaddition, such administration also encompasses use of each type oftherapeutic agent in a sequential or separate manner, either atapproximately the same time or at different times. Regardless of whetherthe active ingredients are administered as a single formulation or inseparate formulations, the drugs are administered to the same patient aspart of the same course of therapy. In any case, the treatment regimenwill provide beneficial effects in treating the conditions or disordersdescribed herein.

An “oral dosage form” includes a unit dosage form prescribed or intendedfor oral administration. In an embodiment of the pharmaceuticalcombinations provided herein, the HDAC6 inhibitor (e.g., Compounds A orB) is administered as an oral dosage form.

The term “HDAC” refers to histone deacetylases, which are enzymes thatremove the acetyl groups from the lysine residues in core histones, thusleading to the formation of a condensed and transcriptionally silencedchromatin. There are currently 18 known histone deacetylases, which areclassified into four groups. Class I HDACs, which include HDAC1, HDAC2,HDAC3, and HDAC8, are related to the yeast RPD3 gene. Class II HDACs,which include HDAC4, HDACS, HDAC6, HDAC7, HDAC9, and HDAC10, are relatedto the yeast Hda1 gene. Class III HDACs, which are also known as thesirtuins are related to the Sir2 gene and include SIRT1-7. Class IVHDACs, which contains only HDAC11, has features of both Class I and IIHDACs. The term “HDAC” refers to any one or more of the 18 known histonedeacetylases, unless otherwise specified.

As used herein, the term “alkyl,” by itself or as part of anothersubstituent means, unless otherwise stated, a straight or branched chainhydrocarbon having the number of carbon atoms designated (i.e.,C₁-C₆-alkyl means an alkyl having one to six carbon atoms) and includesstraight and branched chains. In an embodiment, C₁-C₆alkyl groups areprovided herein. Examples include methyl, ethyl, propyl, isopropyl,butyl, isobutyl, tert-butyl, pentyl, neopentyl, and hexyl. Otherexamples of C₁-C₆-alkyl include ethyl, methyl, isopropyl, isobutyl,n-pentyl, and n-hexyl.

As used herein, the term “alkoxy,” refers to the group —O-alkyl, whereinalkyl is as defined herein. Alkoxy includes, by way of example, methoxy,ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, t-butoxy and thelike. In an embodiment, C₁-C₆ alkoxy groups are provided herein.

As used herein, the term “halo” or “halogen” alone or as part of anothersubstituent means, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom, preferably, fluorine, chlorine, or bromine,more preferably, fluorine or chlorine.

As used herein, the term “cycloalkyl” means a non-aromatic carbocyclicsystem that is partially or fully saturated having 1, 2 or 3 ringswherein such rings may be fused. The term “fused” means that a secondring is present (i.e., attached or formed) by having two adjacent atomsin common (i.e., shared) with the first ring. Cycloalkyl also includesbicyclic structures that may be bridged or spirocyclic in nature witheach individual ring within the bicycle varying from 3-8 atoms. The term“cycloalkyl” includes, but is not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, bicyclo[3.1.0]hexyl, spiro[3.3]heptanyl, andbicyclo[1.1.1]pentyl. In an embodiment, C₄-C₇ cycloalkyl groups areprovided herein.

As used herein, the term “heterocycloalkyl” means a non-aromaticcarbocyclic system containing 1, 2, 3 or 4 heteroatoms selectedindependently from N, O, and S and having 1, 2 or 3 rings wherein suchrings may be fused, wherein fused is defined above. Heterocycloalkylalso includes bicyclic structures that may be bridged or spirocyclic innature with each individual ring within the bicycle varying from 3-8atoms, and containing 0, 1, or 2 N, O, or S atoms. The term“heterocycloalkyl” includes cyclic esters (i.e., lactones) and cyclicamides (i.e., lactams) and also specifically includes, but is notlimited to, epoxidyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl(i.e., oxanyl), pyranyl, dioxanyl, aziridinyl, azetidinyl, pyrrolidinyl,2,5-dihydro-1H-pyrrolyl, oxazolidinyl, thiazolidinyl, piperidinyl,morpholinyl, piperazinyl, thiomorpholinyl, 1,3-oxazinanyl,1,3-thiazinanyl, 2-azabicyclo[2.1.1]hexanyl, 5-azabicyclo[2.1.1]hexanyl,6-azabicyclo[3.1.1] heptanyl, 2-azabicyclo[2.2.1]heptanyl,3-azabicyclo[3.1.1]heptanyl, 2-azabicyclo[3.1.1]heptanyl,3-azabicyclo[3.1.0]hexanyl, 2-azabicyclo[3.1.0]hexanyl,3-azabicyclo[3.2.1]octanyl, 8-azabicyclo[3.2.1]octanyl,3-oxa-7-azabicyclo[3.3.1]nonanyl, 3-oxa-9-azabicyclo[3.3.1]nonanyl,2-oxa-5-azabicyclo[2.2.1]heptanyl, 6-oxa-3-azabicyclo[3.1.1]heptanyl,2-azaspiro[3.3]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl,2-oxaspiro[3.3]heptanyl, 2-oxaspiro[3.5]nonanyl, 3-oxaspiro[5.3]nonanyl,and 8-oxabicyclo[3.2.1]octanyl. In an embodiment, C₂-C₇ heterocycloalkylgroups are provided herein.

As used herein, the term “aromatic” refers to a carbocycle orheterocycle with one or more polyunsaturated rings and having aromaticcharacter, i.e., having (4n+2) delocalized π (pi) electrons, where n isan integer.

As used herein, the term “aryl” means an aromatic carbocyclic systemcontaining 1, 2 or 3 rings, wherein such rings may be fused, whereinfused is defined above. If the rings are fused, one of the rings must befully unsaturated and the fused ring(s) may be fully saturated,partially unsaturated or fully unsaturated. The term “aryl” includes,but is not limited to, phenyl, naphthyl, indanyl, and1,2,3,4-tetrahydronaphthalenyl. In some embodiments, aryl groups have 6carbon atoms. In some embodiments, aryl groups have from six to tencarbon atoms. In some embodiments, aryl groups have from six to sixteencarbon atoms. In an embodiment, C₅-C₇ aryl groups are provided herein.

As used herein, the term “heteroaryl” means an aromatic carbocyclicsystem containing 1, 2, 3, or 4 heteroatoms selected independently fromN, O, and S and having 1, 2, or 3 rings wherein such rings may be fused,wherein fused is defined above. The term “heteroaryl” includes, but isnot limited to, furanyl, thiophenyl, oxazolyl, thiazolyl, imidazolyl,pyrazolyl, triazolyl, tetrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl,thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,imidazo[1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl,5,6,7,8-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydroquinolinyl,6,7-dihydro-5H-cyclopenta[b]pyridinyl,6,7-dihydro-5H-cyclopenta[c]pyridinyl,1,4,5,6-tetrahydrocyclopenta[c]pyrazolyl,2,4,5,6-tetrahydrocyclopenta[c]pyrazolyl,5,6-dihydro-4H-pyrrolo[1,2-b]pyrazolyl,6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazolyl,5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyridinyl,4,5,6,7-tetrahydropyrazolo[1,5-a]pyridinyl,4,5,6,7-tetrahydro-1H-indazolyl and 4,5,6,7-tetrahydro-2H-indazolyl. Inan embodiment, C₂-C₇ heteroaryl groups are provided herein.

As used herein, the term “polycyclic ring” means two or more carbocyclicrings in which two or more carbons are common to two adjoining rings(the rings are “fused rings”) wherein at least one of the rings isaromatic, e.g., the other cyclic rings may be cycloalkyls, and/or aryls.The term “polycyclic ring” includes, but is not limited to, fluorene,anthracene, 9,10-dihydroanthracene, phenanthrene,9,10-dihydrophenanthrene, phenalene, and 2,3-dihydrophenalene. In anembodiment, C₉-C₁₆ polycyclic rings are provided herein.

As used herein, the term “bicyclic heteroaryl” means 7 to 11 memberedbicyclic aromatic groups which have at least one heteroatom in at leastone carbon atom-containing ring. Each ring of the heteroaryl groupcontaining a heteroatom may have 1, 2, 3, or 4 heteroatoms selected fromnitrogen atoms, oxygen atoms and/or sulfur atoms, where the nitrogenand/or sulfur heteroatoms may optionally be oxidized and the nitrogenheteroatoms may optionally be quaternized. Exemplary bicyclic heteroarylgroups include benzothiazolyl, benzoxazolyl, benzoxadiazolyl,benzothienyl, quinolinyl, chromenyl, indolyl, indazolyl, isoquinolinyl,benzimidazolyl, benzopyranyl, benzofuryl, benzofurazanyl, benzopyranyl,cinnolinyl, quinoxalinyl, pyrrolopyridyl, furopyridinyl (such asfuro[2,3-c]pyridinyl, furo[3,2-b]pyridinyl] or furo[2,3-b]pyridinyl),triazinylazepinyl, and the like. In an embodiment, C₅-C₁₅ biciclicheteroaryl groups are provided herein.

It is to be understood that if an aryl, heteroaryl, cycloalkyl, orheterocycloalkyl moiety may be bonded or otherwise attached to adesignated moiety through differing ring atoms (i.e., shown or describedwithout denotation of a specific point of attachment), then all possiblepoints are intended, whether through a carbon atom or, for example, atrivalent nitrogen atom. For example, the term “pyridinyl” means 2-, 3-or 4-pyridinyl, the term “thiophenyl” means 2- or 3-thiophenyl, and soforth.

As used herein, the term “substituted” means that an atom or group ofatoms has replaced hydrogen as the substituent attached to anothergroup.

Compounds of the Invention

In an embodiment, the invention provides a compound of Formula I:

or a pharmaceutically acceptable salt thereof;

wherein

R¹ is H, furan, pyridine, phenyl or thiophene, wherein phenyl isoptionally substituted with halogen;

R² is H or C₁₋₄ alkyl, wherein the C₁₋₄ alkyl group is optionally,independently substituted one or more times with halo, —CN, —NO₂, —C₁-C₆alkoxy, —C₁-C₆alkyl, —C₁-C₆ haloalkyl, and C(O)—C₁-C₆ alkyl; and

B is oxazole, phenyl, pyridine, thiophene, thiazole or quinoline,wherein the thiazole is optionally substituted with CH₃, and wherein ifR¹ is thiophene, B is not 2-(piperazin-1-yl)quinoline.

In another embodiment of Formula I, R¹ is H, furan, pyridine, phenyl orthiophene, wherein phenyl is optionally substituted with halogen;

R² is H or CH₃; and

B is phenyl, pyridine, thiophene, thiazole or quinoline, wherein thethiazole is optionally substituted with CH₃, and wherein if R¹ isthiophene, B is not 2-(piperazin-1-yl)quinoline.

In further embodiment of Formula I, R¹ is furan, pyridine, phenyl orthiophene, wherein phenyl is optionally substituted with halogen.

In a specific embodiment, including pharmaceutically acceptable saltsthereof, Formula I is:

In an embodiment, the invention provides a compound of Formula II:

or a pharmaceutically acceptable salt thereof;

wherein

R³ is furan, pyridine, phenyl or thiophene, wherein phenyl is optionallysubstituted with halogen;

R⁴ is H or C₁₋₄ alkyl, wherein the C₁₋₄ alkyl group is optionally,independently substituted one or more times with halo, —CN, —NO₂, —C₁-C₆alkoxy, —C₁-C₆alkyl, —C₁-C₆ haloalkyl, or C(O)—C₁-C₆ alkyl; and

B is pyrazine, oxazole, pyridine, thiophene, or thiazole.

In another embodiment of Formula II, R³ is furan, pyridine, phenyl orthiophene, wherein

phenyl is optionally substituted with halogen;

R⁴ is H or CH₃; and

B is pyrazine, oxazole, pyridine, thiophene, or thiazole.

In another embodiment of Formula II, R³ is furan, pyridine, orthiophene;

R⁴ is H or C₁-C₄ alkyl; and

B is a five-membered heteroaryl ring.

In another embodiment of Formula II, R³ is furan, pyridine, orthiophene;

R⁴ is H or C₁-C₄ alkyl; and

B is oxazole, thiophene, or thiazole.

In another embodiment of Formula II, R³ is furan or thiophene;

R⁴ is H or C₁-C₄ alkyl; and

B is thiophene or thiazole.

In a specific embodiment, including pharmaceutically acceptable saltsthereof, Formula II is:

In an embodiment, the invention provides a compound of Formula III:

or a pharmaceutically acceptable salt thereof;

wherein

(a) X is C(H), Y is C(H), and Z is N;

R⁵ is H or phenyl; and

R⁶ is H; or

(b) X is C(H), Y is C(H), and Z is N;

R⁵ is 1-furan; and

R⁶ is C₁₋₄ alkyl, wherein the C₁₋₄ alkyl group is optionally,independently substituted one or more times with halo, —CN, —NO₂, —C₁-C₆alkoxy, —C₁-C₆ alkyl, —C₁-C₆ haloalkyl, and C(O)—C₁-C₆ alkyl; or

(c) X is C(H), Y is N, and Z is C(H);

R⁵ is H, 1-furan, 2-furan, pyridine, or thiophene; and

R⁶ is H; or

(d) X is N, Y is C(H), and Z is C(H);

R⁵ is H, 1-furan, 2-furan, pyridine, or phenyl, wherein phenyl isoptionally substituted with halogen; and

R⁶ is H.

In another embodiment of Formula III, X is C(H), Y is C(H), and Z is N;R⁵ is 1-furan; and R⁶ is CH₃.

In another embodiment of Formula III, X is C(H), Y is C(H), and Z is N;R⁵ is H; and R⁶ is H.

In a specific embodiment, including pharmaceutically acceptable saltsthereof, Formula III is:

In an embodiment, the invention provides a compound of Formula IV:

or a pharmaceutically acceptable salt thereof;

wherein

R⁷ is H, furan, pyridine, or thiophene.

In a specific embodiment, including pharmaceutically acceptable saltsthereof, Formula IV is:

Certain embodiments of Formula I, II, III, or IV, or pharmaceuticallyacceptable salts thereof, are shown below in Table 1. Compounds ofFormula I, II, III, or IV, or pharmaceutically acceptable salts thereof,and compounds of Table 1, or pharmaceutically acceptable salts thereof,are sometimes referred to herein as “compounds of the invention,” or“compounds provided herein.”

TABLE 1 Com- pound Structure No.

001

002

003

004

005

006

007

008

009

010

011

012

013

014

015

016

017

018

019

020

021

022

023

024

025

026

027

028

029

030

031

032

033

034

035

036

037

038

039

040

041

042

043

044

045

046

047

048

049

050

051

052

053

054

055

056

057

058

059

060

061

062

063

064

065

066

067

068

069

070

071

072

073

074

075

076

077

078

079

080

081

082

083

084

085

086

087

088

089

090

In one embodiment, the disclosed compounds may exist as tautomers. Alltautomers are included within the scope of the compounds presentedherein.

Compounds described herein also include isotopically-labeled compoundswherein one or more atoms is replaced by an atom having the same atomicnumber, but an atomic mass or mass number different from the atomic massor mass number usually found in nature. Examples of isotopes suitablefor inclusion in the compounds described herein include and are notlimited to ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ³⁶Cl, ¹⁸F, ¹²³I, ¹²⁵I, ¹³N, ¹⁵N, ¹⁵O,¹⁷O, ¹⁸O, ³²P, and ³⁵S. In one embodiment, isotopically-labeledcompounds are useful in drug or substrate tissue distribution studies.In another embodiment, substitution with heavier isotopes such asdeuterium affords greater metabolic stability (for example, increased invivo half-life or reduced dosage requirements). In another embodiment,the compounds described herein include a ²H (i.e., deuterium) isotope.

In yet another embodiment, substitution with positron emitting isotopes,such as ¹¹C, ¹⁸F, ¹⁵O and ¹³N, is useful in Positron Emission Topography(PET) studies for examining substrate receptor occupancy.Isotopically-labeled compounds are prepared by any suitable method or byprocesses using an appropriate isotopically-labeled reagent in place ofthe non-labeled reagent otherwise employed.

The specific compounds described herein, and other compounds encompassedby one or more of the Formulas described herein having differentsubstituents are synthesized using techniques and materials describedherein and as described, for example, in Fieser and Fieser's Reagentsfor Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd'sChemistry of Carbon Compounds, Volumes 1-5 and Supplementals (ElsevierScience Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wileyand Sons, 1991), Larock's Comprehensive Organic Transformations (VCHPublishers Inc., 1989), March, Advanced Organic Chemistry 4^(th) Ed.,(Wiley 1992); Carey and Sundberg, Advanced Organic Chemistry 4th Ed.,Vols. A and B (Plenum 2000, 2001), and Green and Wuts, Protective Groupsin Organic Synthesis 3rd Ed., (Wiley 1999) (all of which areincorporated by reference for such disclosure). General methods for thepreparation of compounds as described herein are modified by the use ofappropriate reagents and conditions, for the introduction of the variousmoieties found in the Formulas as provided herein.

Compounds described herein are synthesized using any suitable proceduresstarting from compounds that are available from commercial sources, orare prepared using procedures described herein.

Methods of Treatment

The compounds of the invention can be used in a method of treating adisease or condition in a subject, said method comprising administeringto the subject a compound of the invention, or a pharmaceuticalcomposition comprising a compound of the invention.

In one aspect, the invention provides a method of selectively inhibitingHDAC1 and/or HDAC2 over other HDACs (e.g., HDAC3 and HDAC6) in asubject, comprising administering to the subject a compound of Formula Ior any of the compounds of Table 1 or pharmaceutically acceptable saltsthereof.

In another aspect, the invention provides a method of selectivelyinhibiting HDAC1 and/or HDAC2 over other HDACs (e.g., HDAC3 and HDAC6)in a subject, comprising administering to the subject a compound ofFormula II, III, IV, or any of the compounds of Table 1 orpharmaceutically acceptable salts thereof.

In one embodiment, the compound of any of the formulae herein (e.g.,Formula I, II, III, or IV) has a selectivity for HDAC1 and/or HDAC2 of 5to 1000 fold over other HDACs.

In another embodiment, the compound of any of the formulae herein (e.g.,Formula I, II, III, or IV) has a selectivity for HDAC1 and/or HDAC2 whentested in a HDAC enzyme assay of about 5 to 1000 fold over other HDACs.

In certain embodiments, the compound has a selectivity for HDAC1 and/orHDAC2 of 15 to 40 fold over other HDACs.

In another aspect, the invention provides a method of treating a diseasemediated by HDAC1 and/or HDAC2 in a subject comprising administering tothe subject a compound of Formula I or any of the compounds of Table 1.

In another aspect, the invention provides a method of treating a diseasemediated by HDAC1 and/or HDAC2 in a subject comprising administering tothe subject a compound of Formula II, III, IV or any of the compounds ofTable 1.

In certain embodiments, the compounds are able to treat a subjectsuffering from or susceptible to a hemoglobinopathy. In a preferredembodiment, the compounds are able to treat sickle-cell disease orbeta-thalessemia.

In another embodiment, the compounds of the invention are useful in thetreatment of myelodysplastic syndromes.

In certain embodiments, the disease is cancer.

In certain embodiments, the cancer is lung cancer, colon and rectalcancer, breast cancer, prostate cancer, liver cancer, pancreatic cancer,brain cancer, kidney cancer, ovarian cancer, stomach cancer, skincancer, bone cancer, gastric cancer, breast cancer, glioma,glioblastoma, neuroblastoma, hepatocellular carcinoma, papillary renalcarcinoma, head and neck squamous cell carcinoma, leukemia, lymphomas,myelomas, retinoblastoma, cervical cancer, melanoma and/or skin cancer,bladder cancer, uterine cancer, testicular cancer, esophageal cancer,and solid tumors. In some embodiments, the cancer is lung cancer, coloncancer, breast cancer, neuroblastoma, leukemia, or lymphomas. In otherembodiments, the cancer is lung cancer, colon cancer, breast cancer,neuroblastoma, leukemia, or lymphoma. In a further embodiment, thecancer is non-small cell lung cancer (NSCLC) or small cell lung cancer.

In a particular embodiment, the cancer is neuroblastoma.

In further embodiments, the cancer is a hematologic cancer, such asleukemia or lymphoma. In a certain embodiment, lymphoma is Hodgkin'slymphoma or Non-Hodgkin's lymphoma. In certain embodiments, leukemia ismyeloid, lymphocytic, myelocytic, lymphoblastic, or megakaryoticleukemia.

In a particular embodiment, the leukemia is acute myelogenous leukemiaand megakaryocytic leukemia.

In another aspect, provided herein is a method for treating sickle celldisease, beta thalassemia, myelodysplastic syndrome, acute myelogenousleukemia, neuroblastoma, or megakaryocytic leukemia in a subjectcomprising administering to the subject in need thereof atherapeutically effective amount of a compound of Formula I, a compoundof Table 1, or a pharmaceutically acceptable salt thereof.

In another aspect, provided herein is a method for treating sickle celldisease, beta thalassemia, myelodysplastic syndrome, acute myelogenousleukemia, neuroblastoma, or megakaryocytic leukemia in a subjectcomprising administering to the subject in need thereof atherapeutically effective amount of a compound of Formula II, III, IV, acompound of Table 1, or a pharmaceutically acceptable salt thereof.

Thus, in another aspect of the invention, methods for the treatment of adisease mediated by HDAC1 and/or HDAC2 are provided comprisingadministering a therapeutically effective amount of a compound ofFormula I, as described herein, to a subject in need thereof. In certainembodiments, the subject is identified as in need of such treatment. Incertain embodiments, a method for the treatment of a diseases isprovided comprising administering a therapeutically effective amount ofa compound of Formula I, or a pharmaceutical composition comprising acompound of Formula I to a subject in need thereof, in such amounts andfor such time as is necessary to achieve the desired result.

In certain embodiments, the method involves the administration of atherapeutically effective amount of a compound of Formula I or apharmaceutically acceptable derivative thereof to a subject (including,but not limited to a human or animal) in need of it (including a subjectidentified as in need).

Thus, in a further aspect of the invention, methods for the treatment ofa disease mediated by HDAC1 and/or HDAC2 are provided comprisingadministering a therapeutically effective amount of a compound ofFormula I, as described herein, to a subject in need thereof. In certainembodiments, the subject is identified as in need of such treatment. Incertain embodiments, a method for the treatment of a disease is providedcomprising administering a therapeutically effective amount of acompound of Formula II, III, or IV, or a pharmaceutical compositioncomprising a compound of Formula II, III, or IV to a subject in needthereof, in such amounts and for such time as is necessary to achievethe desired result.

In certain embodiments, the method involves the administration of atherapeutically effective amount of a compound of Formula II, III, orIV, or a pharmaceutically acceptable derivative thereof to a subject(including, but not limited to a human or animal) in need of it(including a subject identified as in need).

Administration/Dosage/Formulations

In another aspect, provided herein is a pharmaceutical compositioncomprising at least one compound of the invention, together with apharmaceutically acceptable carrier.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

In particular, the selected dosage level will depend upon a variety offactors including the activity of the particular compound employed, thetime of administration, the rate of excretion of the compound, theduration of the treatment, other drugs, compounds or materials used incombination with the compound, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well, known in the medical arts.

A medical doctor, e.g., physician or veterinarian, having ordinary skillin the art may readily determine and prescribe the effective amount ofthe pharmaceutical composition required. For example, the physician orveterinarian could begin administration of the pharmaceuticalcomposition to dose the disclosed compound at levels lower than thatrequired in order to achieve the desired therapeutic effect andgradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulatethe compound in dosage unit form for ease of administration anduniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the patients tobe treated; each unit containing a predetermined quantity of thedisclosed compound calculated to produce the desired therapeutic effectin association with the required pharmaceutical vehicle. The dosage unitforms of the invention are dictated by and directly dependent on (a) theunique characteristics of the disclosed compound and the particulartherapeutic effect to be achieved, and (b) the limitations inherent inthe art of compounding/formulating such a disclosed compound for thetreatment of pain, a depressive disorder, or drug addiction in apatient.

In one embodiment, the compounds of the invention are formulated usingone or more pharmaceutically acceptable excipients or carriers. In oneembodiment, the pharmaceutical compositions of the invention comprise atherapeutically effective amount of a disclosed compound and apharmaceutically acceptable carrier.

Routes of administration of any of the compositions of the inventioninclude oral, nasal, rectal, intravaginal, parenteral, buccal,sublingual or topical. The compounds for use in the invention may beformulated for administration by any suitable route, such as for oral orparenteral, for example, transdermal, transmucosal (e.g., sublingual,lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- andperivaginally), (intra)nasal and (trans)rectal), intravesical,intrapulmonary, intraduodenal, intragastrical, intrathecal,subcutaneous, intramuscular, intradermal, intra-arterial, intravenous,intrabronchial, inhalation, and topical administration. In oneembodiment, the preferred route of administration is oral.

Suitable compositions and dosage forms include, for example, tablets,capsules, caplets, pills, gel caps, troches, dispersions, suspensions,solutions, syrups, granules, beads, transdermal patches, gels, powders,pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs,suppositories, liquid sprays for nasal or oral administration, drypowder or aerosolized formulations for inhalation, compositions andformulations for intravesical administration and the like. It should beunderstood that the formulations and compositions that would be usefulin the present invention are not limited to the particular formulationsand compositions that are described herein.

For oral application, particularly suitable are tablets, dragees,liquids, drops, suppositories, or capsules, caplets and gel caps. Thecompositions intended for oral use may be prepared according to anymethod known in the art and such compositions may contain one or moreagents selected from the group consisting of inert, non-toxicpharmaceutically excipients that are suitable for the manufacture oftablets. Such excipients include, for example an inert diluent such aslactose; granulating and disintegrating agents such as cornstarch;binding agents such as starch; and lubricating agents such as magnesiumstearate. The tablets may be uncoated or they may be coated by knowntechniques for elegance or to delay the release of the activeingredients. Formulations for oral use may also be presented as hardgelatin capsules wherein the active ingredient is mixed with an inertdiluent.

For parenteral administration, the disclosed compounds may be formulatedfor injection or infusion, for example, intravenous, intramuscular orsubcutaneous injection or infusion, or for administration in a bolusdose or continuous infusion. Suspensions, solutions or emulsions in anoily or aqueous vehicle, optionally containing other formulatory agentssuch as suspending, stabilizing or dispersing agents may be used.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures, embodiments, claims, and examples described herein.Such equivalents were considered to be within the scope of thisinvention and covered by the claims appended hereto. For example, itshould be understood, that modifications in reaction conditions,including but not limited to reaction times, reaction size/volume, andexperimental reagents, such as solvents, catalysts, pressures,atmospheric conditions, e.g., nitrogen atmosphere, andreducing/oxidizing agents, with art-recognized alternatives and using nomore than routine experimentation, are within the scope of the presentapplication.

It is to be understood that wherever values and ranges are providedherein, all values and ranges encompassed by these values and ranges,are meant to be encompassed within the scope of the present invention.Moreover, all values that fall within these ranges, as well as the upperor lower limits of a range of values, are also contemplated by thepresent application.

The following examples further illustrate aspects of the presentinvention. However, they are in no way a limitation of the teachings ordisclosure of the present invention as set forth.

EXAMPLES

The invention is further illustrated by the following examples, whichshould not be construed as further limiting. The practice of the presentinvention will employ, unless otherwise indicated, conventionaltechniques of organic synthesis, cell biology, cell culture, molecularbiology, transgenic biology, microbiology and immunology, which arewithin the skill of the art.

Abbreviations

-   -   Ac acetyl    -   QC degree Celsius    -   dba dibenzylideneacetone    -   DCM dichloromethane    -   DIPEA N,N-Diisopropylethylamine    -   DMAP 4-dimethylaminopyridine    -   DMF dimethylformamide    -   DMSO dimethylsulfoxide    -   dppf 1,1′-bis(diphenylphosphino)ferrocene    -   EDCI 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimidhydrochloride    -   EA ethyl acetate    -   HATU 0-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium        Hexafluorophosphate    -   HOAt 1-hydroxy-7-azabenzotriazole    -   HOBt 1-hydroxybenzotriazole    -   HPLC high performance liquid chromatography    -   m-CPBA m-chloroperoxybenzoic acid    -   PE petroleum ether    -   Ph phenyl    -   Py pyridine    -   Ruphos 2-Dicyclohexylphosphino-2′,6′-di-i-propoxy-1,1′-biphenyl    -   S-Phos 2-Dicyclohexylphosphino-2′,6′-dimethoxybiphenyl    -   TFA trifluoroacetic acid    -   THF tetrahydrofuran    -   TLC thin layer chromatography    -   Tol toluene or tolyl    -   Xantphos 4,5-Bis(diphenylphosphino)-9,9-dimethylxanthene

Synthesis Procedures Example 1—Synthesis of Compound 001

Step 1:

To a solution of methyl quinoline-6-carboxylate (3.6 g, 20 mmol) in DCM(60 mL) was slowly added m-CPBA (9.0 g, 40 mmol) at room temperature,the mixture was stirred at room temperature overnight. After completion,the mixture was quenched by aqueous saturated NaHCO₃, then washed withwater, concentrated to get a residue which was purified byrecrystallization from EtOH added DCM (200 mL), added aqueous saturatedNaHCO₃ at 0° C. to PH>7. The organic layer was separated, dried,filtered to afford compound 2 as a yellow solid (4.6 g, 77%).

Step 2:

To a solution of 6-(methoxycarbonyl)quinoline 1-oxide (2.0 g, 10 mmol)in Ac₂O (50 mL), the mixture was heated at 135° C. overnight. Aftercompletion, the mixture was diluted with water, extracted with EA,washed with saturated NaHCO₃, purified on column(DCM/MeOH/NH3-H2O=250:50:1) to afford the crude. The mixture waspurified by recrystallization from EA to afford compound 3 as a palesolid (1.4 g, 70%).

Step 3:

To a solution of compound methyl 2-hydroxyquinoline-6-carboxylate (10 g,0.049 mol) and DMF (20 mL) in DCM (100 mL) was slowly added SOCl₂ (20mL) at 0° C., the mixture was stirred at room temperature for 5 hours.TLC monitored completion, then the mixture was concentrated to get aresidue, added DCM (200 mL), added aqueous Sat. NaHCO₃ at 0° C. to PH>7.The organic layer was separated, dried, filtered (flash silica gel) andconcentrated to afford compound 4 (10 g, 92%) as a yellow solid.

Step 4:

A mixture of compound methyl 2-chloroquinoline-6-carboxylate (10 g,0.045 mol), CuI (10 g, 0.53 mol), N-boc-piperazine (25 g, 0.135 mol) andK₂CO₃ (18.6 g, 0.135 mol) was stirred at 100° C. for overnight. TLCmonitored completion. The mixture was added EA (300 mL), filtered (flashsilica gel), concentrated to get a residue, added water (300 mL) andaqueous saturated citric acid (30 mL), stirred at room temperature for30 minutes, filtered to obtain compound 5 (18 g, crude) as yellow solidwithout the further purification.

Step 5:

A mixture of compound methyl2-(4-(tert-butoxycarbonyl)piperazin-1-yl)quinoline-6-carboxylate (18 g,crude) and 2M NaOH (50 mL) in EtOH (100 mL) and THF (100 mL) was stirred70° C. for 4 hours. TLC monitored completion. The mixture concentratedto a residue, added water (300 mL) and aqueous saturated citric acid (40mL), filtered to obtain compound 6 (14.5 g, 91% for two step) as ayellow solid.

Step 6:

A mixture of compound 6 (357 mg, 1 mmol), benzene-1,2-diamine (100 mg,0.9 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (380 mg, 2 mmol) in pyridine (10 mL) was stirred at roomtemperature for overnight. After completed, the mixture wasconcentrated, poured into water (100 mL), filtered to obtain compound 7(350 mg, crude) as a yellow solid.

Step 7:

A mixture of compound 7 (350 mg, crude) and TFA (3 mL) in DCM (3 mL) wasstirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 001 (119 mg, 34%, 2 steps) as yellow solid.LCMS: m/z=348 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 9.94 (s, 1H), 8.18 (dd,J=9.0, 2.0 Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 7.63 (dd, J=16.5, 1.5 Hz,2H), 7.41 (m, 2H), 7.28-6.94 m, 4H), 4.00 (m, 4H), 3.27 (s, 4H).

Example 2—Synthesis of Compound 002

Step 1:

A mixture of compound 1 (357 mg, 1 mmol), tert-butyl(4-amino-[1,1′-biphenyl]-3-yl)carbamate (260 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (380 mg, 2mmol) in pyridine (12 mL) was stirred at room temperature for overnight.After completed, the mixture was concentrated, poured into water (80mL), filtered to get compound 2 (415 mg, crude) as a yellow solid.

Step 2:

A mixture of compound 2 (415 mg, crude) and TFA (4 mL) in DCM (4 mL) wasstirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 002 (139 mg, 33%, 2 steps) as yellow solid.LCMS: m/z=424 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 9.94 (s, 1H), 8.99 (brs,2H), 8.49 (d, J=1.5 Hz, 1H), 8.27 (d, J=9.5 Hz, 1H), 8.18 (dd, J=9.0,2.0 Hz, 1H), 7.70 (d, J=8.5 Hz, 1H), 7.63 (dd, J=16.5, 1.5 Hz, 2H), 7.41(m, 2H), 7.28-6.94 (m, 5H), 4.00 (m, 4H), 3.27 (s, 4H).

Example 3—Synthesis of Compound 003

Step 1:

Under N₂, a mixture of tert-butyl 4-bromo-2-nitrophenylcarbamate (1.00g, 3.15 mmol) and furan-2-ylboronic acid (494 mg, 4.41 mmol),1,1′-Bis(diphenylphosphino)ferrocene-palladium(II)dichloridedichloromethane complex (200 mg, 0.24 mmol) and potassium carbonate (872mg, 6.31 mmol) in dioxane (40 mL) and water (8 mL) was heated at 90° C.for 18 hours. Then it was cooled to room temperature, and dioxane wasremoved in vacuo. The residue was extracted with EA and concentrated invacuo. The residue was purified by silica gel chromatography (PE,PE/EA=50/1) to give compound 2 as a yellow solid (723 mg, 76%).

Step 2:

To a mixture of tert-butyl 4-(furan-2-yl)-2-nitrophenylcarbamate (673mg, 2.21 mmol), FeCl₃ (219 mg, 5.21 mmol) and active charcoal (200 mg)in EtOH (30 mL) was added N₂H₄ (4 mL). It was stirred at 60° C. for 2 h.Then it was filtrated and concentrated in vacuo. And the residue waspartitioned with EA and water. The combined EA layers were concentratedin vacuo and the residue was purified by silica gel chromatography(PE/EA=5/1, DCM) to give compound 3 as a white solid (576 mg, 95%).LCMS: m/z=219.0 (M-56+H)⁺.

Step 3:

A mixture of2-(4-(tert-butoxycarbonyl)piperazin-1-yl)quinoline-6-carboxylic acid(140 mg, 0.39 mmol) and HATU (140 mg, 0.43 mmol) in DMF (10 mL) wasstirred at room temperature for 15 minutes. Then tert-butyl2-amino-4-(furan-2-yl)phenylcarbamate (108 mg, 0.43 mmol) was added. Itwas stirred at 50° C. for 18 hours. Then it was poured into water. Themixture was extracted with EA. The combined EA layers were concentratedin vacuo and the residue was purified by prep-TLC to give the compound 4(261 mg). LCMS: m/z=614.2 (M+H)⁺.

Step 4:

To a mixture of tert-butyl3-(5-(4-methylpiperazin-1-yl)picolinamido)biphenyl-4-ylcarbamate (261mg) in MeOH (2 mL) was added HCl/dioxane (4 mL) at 0° C. It was stirredat 0° C. to room temperature for 18 h. It was concentrated in vacuo andthe residue was purified by prep-HPLC to give compound 003 as a whitesolid (56 mg, lot SP-0017146-043). LCMS: m/z=414.2 (M+H)⁺. ¹H NMR (500MHz, DMSO) δ 9.94 (s, 1H), 8.99 (brs, 2H), 8.49 (d, J=1.5 Hz, 1H), 8.27(d, J=9.5 Hz, 1H), 8.18 (dd, J=9.0, 2.0 Hz, 1H), 7.70 (d, J=8.5 Hz, 1H),7.63 (dd, J=16.5, 1.5 Hz, 2H), 7.41 (m, 2H), 6.94 (d, J=8.5 Hz, 1H),6.68 (d, J=3.2 Hz, 1H), 6.53 (m, 1H), 4.00 (m, 4H), 3.27 (s, 4H).

Example 4—Synthesis of Compound 004

Step 1:

To a solution of compound 1 (3.16 g, 10 mmol) in dioxane (20 mL) wasadded furan-3-ylboronic acid (1.45 g, 13 mmol), K₃PO₄ (4.23 g, 20 mmol)and Pd[P(C₆H₅)₃]₄ (700 mg, 0.7 mmol). The mixture was stirred at 90° C.for overnight. After completed, water was added and the mixture wasextracted with EA (2×150 mL), the organic layer was purified by silicacolumn chromatography (PE/EA 2/1) to afford compound 2 (2.2 g, 72%) as ayellow oil.

Step 2:

To a solution of compound 2 (200 mg, 0.65 mmol) in EA (10 mL) was addedPd/C (20 mg). The reaction mixture was stirred under H₂ for 5 hour atroom temperature. After completion, the mixture was filtrated, extractedwith EA (2×50 mL) to afford compound 3 (180 mg, 100%) as a yellow solid.

Step 3:

A mixture of compound 3 (180 mg, 0.66 mmol),2-(4-(tert-butoxycarbonyl)piperazin-1-yl)quinoline-6-carboxylic acid(260 mg, 0.73 mmol), and EDCI (835 mg, 4.4 mmol) in pyridine (5 mL) wasstirred at room temperature for overnight, TLC monitored. Put into waterand extracted with EA (2×100 mL), the organic layer was separated,dried, and purified by prep-TLC (PE/EA 2/1) to afford compound 4 (250mg, 62%) as an off-white solid.

Step 4:

To a solution of compound 4 (250 mg, 0.4 mmol) in DCM (5 mL) was addedTFA (2 mL), the mixture was stirred at room temperature for 2 hours.After completed, the resulting mixture was concentrated and alkalifiedwith (aqueous) NaHCO₃ to pH=8. The precipitate was collected and washedby water to afford compound 004 as a white powder (100 mg, 59%). LCMS:m/z=414 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.77 (s, 1H), 8.41 (d, J=1.7Hz, 1H), 8.11 (dd, J=11.3, 5.4 Hz, 2H), 7.97 (s, 1H), 7.67 (t, J=1.6 Hz,1H), 7.59 (d, J=8.8 Hz, 1H), 7.42 (d, J=1.8 Hz, 1H), 7.33-7.22 (m, 2H),6.81 (d, J=8.2 Hz, 2H), 5.01 (s, 2H), 3.74-3.63 (m, 4H), 2.88-2.73 (m,4H).

Example 5—Synthesis of Compound 005

Step 1:

2-(4-(tert-butoxycarbonyl) piperazin-1-yl) quinoline-6-carboxylic acid(108 mg, 0.3 mmol), tert-butyl 2-amino-4-(pyridin-4-yl) phenylcarbamate(86 mg, 0.3 mmol) and EDCI (173 mg, 0.9 mmol) were added into pyridine(3 ml). The mixture was stirred for overnight at room temperature. Whenthe reaction finished, it was extracted by EA and washed by NaHCO₃ andsaturated brine. Then the EA layer was concentrated to afford compound 2(200 mg, crude).

Step 2:

Tert-butyl 4-(6-(2-(tert-butoxycarbonylamino)-5-(pyridin-4-yl)phenylcarbamoyl) quinolin-2-yl) piperazine-1-carboxylate (260 mg, crude)was added into DCM (2 ml). Then TFA (2 ml) was added. The mixture wasstirred at room temperature for 1 hour. It was purified by Prep-HPLC(base method). Compound 005 was obtained as a white solid (90 mg, 51%yield, lot SP-0017467-057). LCMS: m/z=425.2 (M+H)⁺. ¹H NMR (400 MHz,DMSO) δ 9.79 (s, 1H), 8.52 (d, J=6.1 Hz, 2H), 8.41 (s, 1H), 8.18-8.05(m, 2H), 7.72 (d, J=2.0 Hz, 1H), 7.60 (d, J=6.4 Hz, 3H), 7.52 (dd,J=8.4, 2.1 Hz, 1H), 7.30 (d, J=9.3 Hz, 1H), 6.90 (d, J=8.4 Hz, 1H), 5.41(s, 2H), 3.68 (d, J=4.7 Hz, 4H), 2.88-2.74 (m, 4H).

Example 6—Synthesis of Compound 006

Step 1:

A mixture of methyl 6-chloronicotinate (1.7 g, 10 mmol), tert-butylpiperazine-1-carboxylate (5.58 g, 30 mmol), DIPEA (6.45 g, 50 mmol) in1,4-dioxane (30 mL) was stirred at 95° C. under N₂ atmosphere forovernight. The mixture was cool and EA (100 mL) and water (100 mL) wereadded, stirred for 30 min, the organic layer was separated, dried,concentrated to get a residue, which was washed by PE (100 mL) to obtaincompound 2 (2.9 g, 91%) as a light yellow solid.

Step 2:

A mixture of compound 2 (3.2 g, 10 mmol) and 2M NaOH (50 mL) in THF (50ml) and EtOH (50 mL) was stirred at 60° C. for 3 h. The mixture wasconcentrated to obtain a residue, HCl (2 M) was added to adjust pH 7,filtered to give compound 3 as a white solid (3.0 g, 97%)

Step 3:

A mixture of compound 3 (307 mg, 1 mmol), tert-butyl3-aminobiphenyl-4-ylcarbamate (256 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (573 mg, 3mmol) in pyridine (15 mL) was stirred at room temperature for overnight.The mixture was poured into water (100 mL), filtered to get compound 4(470 mg, 82%) as a yellow solid.

Step 4:

A mixture of compound 4 (286 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 h. The mixture was purified byPrep-HPLC to obtain compound 006 (84 mg, 45%) as a yellow solid. LCMS:m/z=374 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.62 (s, 1H), 8.85 (s, 2H),8.81 (s, 1H), 8.20 (d, J=8.9 Hz, 1H), 7.55 (d, J=7.7 Hz, 2H), 7.50 (s,1H), 7.39 (t, J=7.7 Hz, 2H), 7.33 (dd, J=8.3, 1.9 Hz, 1H), 7.24 (t,J=7.3 Hz, 1H), 7.02 (d, J=9.1 Hz, 1H), 6.87 (d, J=8.3 Hz, 1H), 5.11 (s,2H), 3.90-3.78 (m, 4H), 3.22 (d, J=4.9 Hz, 4H).

Example 7—Synthesis of Compound 007

Step 1:

A mixture of compound 3 (307 mg, 1 mmol), tert-butyl2-amino-4-(thiophen-2-yl)phenyl-carbamate (261 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (573 mg, 3mmol) in pyridine (15 mL) was stirred at room temperature for overnight.The mixture was poured into water (100 mL), filtered to obtain compound4 (463 mg, 80%) as a yellow solid.

Step 2:

A mixture of compound 4 (290 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 007 (95 mg, 50%) as a yellow solid. LCMS:m/z=380 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.62 (s, 1H), 8.90 (s, 2H),8.81 (s, 1H), 8.19 (d, J=7.2 Hz, 1H), 7.45 (s, 1H), 7.36 (d, J=5.0 Hz,1H), 7.31 (d, J=8.4 Hz, 1H), 7.25 (d, J=3.4 Hz, 1H), 7.09-6.98 (m, 2H),6.81 (d, J=8.3 Hz, 1H), 5.20 (s, 2H), 3.86 (s, 4H), 3.22 (s, 4H).

Example 8—Synthesis of Compound 008

Step 1:

To a solution of compound 1 (175 mg, 0.57 mmol) in pyridine (4 ml) wasadded tert-butyl (2-amino-4-(furan-2-yl)phenyl)carbamate (156 mg, 0.57mmol) and EDCI (218 mg). The mixture was stirred at room temperature forovernight. The solution was evaporated off and washed with HCl (1 N),extracted by EA (20 ml×3), washed with water, brine, purified byprep-TLC (PE/EA=1/1) to afford compound 3 as a white solid (128 mg,40%).

Step 2:

To a solution of compound 3 (124 mg, 0.22 mmol) in DCM (3 ml) was addedTFA (1 ml). The mixture was stirred at room temperature for overnight.The solution was concentrated, the pH adjusted to >7, filtered, andwashed with water to afford compound 008 as a yellow solid (77 mg,97.4%). ¹H NMR (400 MHz, DMSO) δ 9.50 (s, 1H), 8.75 (d, J=2.3 Hz, 1H),8.08 (dd, J=9.0, 2.4 Hz, 1H), 7.60 (d, J=1.2 Hz, 1H), 7.51 (d, J=1.8 Hz,1H), 7.31 (dd, J=8.4, 2.0 Hz, 1H), 6.86 (d, J=9.1 Hz, 1H), 6.80 (d,J=8.4 Hz, 1H), 6.60 (d, J=3.3 Hz, 1H), 6.50 (dd, J=3.2, 1.8 Hz, 1H),5.14 (s, 2H), 3.58-3.50 (m, 4H), 2.81-2.72 (m, 4H). LCMS: m/z=364.2(M+H)⁺

Example 9—Synthesis of Compound 009

Step 1:

6-(4-(tert-butoxycarbonyl) piperazin-1-yl) nicotinic acid (92 mg, 0.3mmol), tert-butyl 2-amino-4-(furan-3-yl) phenylcarbamate (82 mg, 0.3mmol) and EDCI (173 mg, 0.9 mmol) were added into pyridine (3 ml). Themixture was stirred at room temperature for overnight. The crudecompound 2 was prepared for next step (200 mg, crude).

Step 2:

Tert-butyl 4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl)phenylcarbamoyl) pyridin-2-yl) piperazine-1-carboxylate (180 mg, crude)was added into DCM (2 ml). Then TFA (2 ml) was added. The mixture wasstirred at room temperature for 6 hours. When the reaction finished, thesolid was filtered and washed by Et₂O. A White solid was affordedcompound 009 (76 mg, 66% yield, lot SP-0017467-083). LCMS: m/z=364.2(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.52 (s, 1H), 8.75 (s, 1H), 8.08 (d,J=8.4 Hz, 1H), 7.97 (s, 1H), 7.66 (s, 1H), 7.37 (s, 1H), 7.23 (d, J=8.1Hz, 1H), 7.04-6.67 (m, 3H), 4.97 (s, 2H), 3.56 (s, 4H), 2.79 (s, 4H).

Example 10—Synthesis of Compound 010

Step 1:

6-(4-(tert-butoxycarbonyl) piperazin-1-yl) nicotinic acid (92 mg, 0.3mmol), tert-butyl 2-amino-4-(pyridin-4-yl) phenylcarbamate (86 mg, 0.3mmol) and EDCI (173 mg, 0.9 mmol) were added into pyridine (3 ml). Themixture was stirred for overnight at room temperature. When the reactionfinished, it was extracted by EA and washed by citric acid, NaHCO₃ andsaturated brine. Then the EA layer was concentrated to afford compound 2as a grey solid (150 mg, 85% yield).

Step 2:

Tert-butyl 4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-4-yl)phenylcarbamoyl) pyridin-2-yl) piperazine-1-carboxylate (150 mg, crude)was added into DCM (2 ml). Then TFA (2 ml) was added. The mixture wasstirred at room temperature for 1 hour. It was purified by Prep-HPLC(base method). Compound 010 was obtained as a light yellow solid (60 mg,61% yield, lot SP-0017467-049). LCMS: m/z=375.1 (M+H)⁺. ¹H NMR (400 MHz,DMSO) δ 9.52 (s, 1H), 8.76 (d, J=2.3 Hz, 1H), 8.51 (dd, J=4.6, 1.5 Hz,2H), 8.09 (dd, J=9.0, 2.4 Hz, 1H), 7.67 (d, J=2.1 Hz, 1H), 7.58 (dd,J=4.7, 1.6 Hz, 2H), 7.50 (dd, J=8.4, 2.2 Hz, 1H), 6.88 (dd, J=8.8, 2.5Hz, 2H), 5.36 (s, 2H), 3.62-0.48 (m, 4H), 2.84-0.73 (m, 4H).

Example 11—Synthesis of Compound 011

Step 1:

A mixture of methyl 4-bromobenzoate (2.15 g, 10 mmol), tert-butylpiperazine-1-carboxylate (5.58 g, 30 mmol), Pd₂(dba)₃ (915 mg, 1 mmol),Xantphos (478 mg, 1 mmol) and Cs₂CO₃ (9.75 g, 30 mmol, 3 eq) in toluene(30 mL) was stirred at 95° C. under N₂ atmosphere for overnight. Themixture was cool and added EA (100 mL), filtered, concentrated to obtaina residue, which was washed by PE (100 mL) to obtain compound 2 (2.88 g,90%) as a light yellow solid.

Step 2:

A mixture of compound 2 (3.2 g, 10 mmol) and 2M NaOH (50 mL) in THF (50ml) and EtOH (50 mL) was stirred at 60° C. for 3 hours. The mixture wasconcentrated to obtain a residue, HCl (2 M) was added to adjust pH to 7,filtered to give compound 3 as a white solid (3.0 g, 100%)

Step 3:

A mixture of compound 3 (306 mg, 1 mmol), tert-butyl3-aminobiphenyl-4-ylcarbamate (256 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (573 mg, 3mmol) in pyridine (15 mL) was stirred at room temperature for overnight.The mixture was poured into water (100 mL) then filtered to obtaincompound 4 (438 mg, 85%) as a yellow solid.

Step 4:

A mixture of compound 4 (286 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 h. The mixture was purified byPrep-HPLC to obtain compound 011 (112 mg, 60%) as a yellow solid. LCMS:m/z=373 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.54 (s, 1H), 7.91 (d, J=8.7Hz, 2H), 7.54 (dd, J=17.4, 4.6 Hz, 3H), 7.40 (t, J=7.6 Hz, 2H), 7.32(dd, J=8.3, 1.8 Hz, 1H), 7.25 (t, J=7.3 Hz, 1H), 7.00 (d, J=8.8 Hz, 2H),6.87 (d, J=8.3 Hz, 1H), 5.06 (s, 2H), 3.25-3.13 (m, 4H), 2.92-2.78 (m,4H).

Example 12—Synthesis of Compound 012

Step 1:

A mixture of compound 3 (306 mg, 1 mmol), tert-butyl2-amino-4-(thiophen-2-yl)phenyl-carbamate (260 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (573 mg, 3mmol) in pyridine (15 mL) was stirred at room temperature for overnight.The mixture was poured into water (100 mL), filtered to obtain compound4 (442 mg, 85%) as a yellow solid.

Step 2:

A mixture of compound 4 (289 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 012 (113 mg, 60%) as a yellow solid. LCMS:m/z=379 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.51 (s, 1H), 7.89 (d, J=8.9Hz, 2H), 7.46 (d, J=2.0 Hz, 1H), 7.36 (dd, J=5.1, 0.8 Hz, 1H), 7.32-7.20(m, 2H), 7.05 (dd, J=5.0, 3.6 Hz, 1H), 6.99 (d, J=8.9 Hz, 2H), 6.81 (d,J=8.3 Hz, 1H), 5.10 (s, 2H), 3.24-3.15 (m, 4H), 2.89-2.77 (m, 4H).

Example 13—Synthesis of Compound 013

Step 1:

A mixture of compound 3 (306 mg, 1 mmol), tert-butyl2-amino-4-(furan-2-yl)phenyl-carbamate (247 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (573 mg, 3mmol) in pyridine (15 mL) was stirred at room temperature for overnight.The mixture was poured into water (100 mL), filtered to obtain compound4 (410 mg, 81%) as a yellow solid.

Step 2:

A mixture of compound 4 (281 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 013 (107 mg, 59%) as a yellow solid. LCMS:m/z=363 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.49 (s, 1H), 7.89 (d, J=8.7Hz, 2H), 7.61 (s, 1H), 7.51 (d, J=1.4 Hz, 1H), 7.32 (d, J=8.3 Hz, 1H),6.99 (d, J=8.7 Hz, 2H), 6.81 (d, J=8.3 Hz, 1H), 6.61 (d, J=3.1 Hz, 1H),6.51 (d, J=1.7 Hz, 1H), 5.10 (s, 2H), 3.20 (s, 4H), 2.84 (s, 4H).

Example 14—Synthesis of Compound 014

Step 1:

A mixture of compound 3 (306 mg, 1 mmol), tert-butyl2-amino-4-(furan-3-yl)-phenylcarbamate (246 mg, 0.9 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (573 mg, 3mmol) in pyridine (15 mL) was stirred at room temperature for overnight.The mixture was poured into water (100 mL), filtered to obtain compound4 (420 mg, 83%) as a yellow solid.

Step 2:

A mixture of compound 4 (281 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 014 (100 mg, 55%) as a yellow solid. LCMS:m/z=363 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.50 (s, 1H), 7.96 (s, 1H),7.89 (d, J=8.7 Hz, 2H), 7.66 (s, 1H), 7.37 (s, 1H), 7.23 (d, J=8.2 Hz,1H), 6.98 (d, J=8.8 Hz, 2H), 6.86-6.73 (m, 2H), 4.92 (s, 2H), 3.19 (d,J=4.8 Hz, 4H), 2.84 (d, J=4.6 Hz, 4H).

Example 15—Synthesis of Compound 015

Step 1:

To a solution of compound 1 (500 mg, 2.32 mmol) in toluene (20 ml) wasadded tert-butyl piperazine-1-carboxylate (1.3 g, 3.0 eq.), Pd₂(dba)₃(213 mg, 0.1 eq.), Ruphos (108 mg, 0.1 eq.) and Cs₂CO₃ (2.273 g, 3.0eq.) at 95° C. overnight under N₂ atmosphere. It was filtered throughcelite and concentrated. It was extracted by EA (2×80 ml). Then it waswashed by Et₂O. Compound 2 was obtained as a yellow solid (623 mg, 84%).

Step 2:

To a solution of compound 2 (623 mg, 1.94 mmol) in THF (8 ml) was added2N NaOH (5 ml) and EtOH (2 ml) at 60° C. overnight. It was concentratedand neutralized by dilute HCl. After filtration, a white solid compound3 (550 mg, 92%) was obtained.

Step 3:

To a solution of compound 4 (3.17 g, 10 mmol) in dioxane (30 ml) and H₂O(6 ml) was added pyridin-4-ylboronic acid (1.6 g, 1.3 eq.), K2CO3 (2.763g, 2.0 eq.), Pd[PPh3]4 (808 mg, 0.07 eq.) at 90° C. overnight under N₂atmosphere. It was filtered with silica gel. It was extracted by EA(2×150 ml) and purified by column chromatography (PE:EA=4:1). A Yellowsolid compound 5 (2.127 g, 67%) was obtained.

Step 4:

To a solution of compound 5 (1.5 g, 4.76 mmol) in EtOH (15 ml) was addedFeCl₃ (154 mg, 0.2 eq.), C (1.5 g) at 60° C. N₂H₄H₂O (2.77 ml, 12.0 eq.)was dropped slowly. It was reacted for 3 hours. It was filtered throughcelite and concentrated. It was washed by H₂O. A brown solid compound 6(895 mg, 66%) was obtained.

Step 5:

A mixture of compound 3 (100 mg, 0.33 mmol), compound 6 (131 mg, 1.4eq.) and EDCI (250 mg, 4.0 eq.) in pyridine (5 ml) were stirred at roomtemperature overnight. It was concentrated and extracted with EA (2×70ml), washed by brine (2×60 ml). It was purified by prep-TLC (PE:EA=1:2).Compound 7 (126 mg, Yield: 67%) was obtained as a white solid.

Step 6:

To a solution of compound 7 (126 mg, 0.22 mmol) in DCM (3 ml) was addedTFA (0.5 ml) at room temperature. The reaction was stirred for 2 hours.Then it was concentrated. It was purified by prep-HPLC. Compound 015 wasobtained as a white solid (26 mg, 32%). LCMS: m/z=374 (M+H)⁺ ¹H NMR (500MHz, DMSO) δ 10.10 (s, 1H), 9.34 (s, 1H), 8.96 (s, 1H), 8.89 (s, 2H),8.06 (d, J=9.2 Hz, 1H), 8.01 (s, 1H), 7.69 (t, J=1.6 Hz, 1H), 7.65 (dd,J=9.3, 2.2 Hz, 1H), 7.46 (s, 1H), 7.40 (d, J=1.9 Hz, 1H), 7.33 (dd,J=8.3, 2.0 Hz, 1H), 6.90 (d, J=8.3 Hz, 1H), 6.84 (s, 1H), 3.70-3.66 (m,4H), 3.32 (s, 4H).

Example 16—Synthesis of Compound 016

Step 1:

Under N₂, a mixture of tert-butyl piperazine-1-carboxylate (1.04 g, 5.55mmol) and methyl 5-bromopicolinate (1.00 g, 4.63 mmol),tris(dibenzylideneacetone)dipalladium (212 mg, 0.23 mmol), S-Phos (384mg, 0.94 mmol) and potassium phosphate (1.38 g, 6.48 mmol) in toluene(20 mL) was heated at 100° C. for 3 days. Then it was cooled to roomtemperature and filtrated. The filtrate was concentrated in vacuo. Andthe residue was purified by silica gel chromatography to give compound 3as a yellow solid (837 mg, 56%).

Step 2:

A mixture of tert-butyl4-(6-(methoxycarbonyl)pyridin-3-yl)piperazine-1-carboxylate (837 mg,2.60 mmol) and LiOH (219 mg, 5.21 mmol) in THF (12 mL), water (6 mL) andMeOH (3 mL) was stirred at 0° C. to room temperature until completion.Then it was concentrated in vacuo to give a crude Li salt of compound 4.LCMS: m/z=308.1 (M+H)+.

Step 3:

A mixture of lithium 5-(4-(tert-butoxycarbonyl)piperazin-1-yl)picolinate(1.30 mmol) and HATU (545 mg, 1.43 mmol) in DMF (4 mL) was stirred atroom temperature for 30 minutes. Then tert-butyl3-aminobiphenyl-4-ylcarbamate (407 mg, 1.43 mmol) was added. It wasstirred at room temperature for 2 h. Then it was poured into water. Theresulting solid was collected by filtration and dried in vacuo to givethe crude compound 5. LCMS: m/z=574.2 (M+H)⁺.

Step 4:

To a mixture of tert-butyl3-(5-(4-methylpiperazin-1-yl)picolinamido)biphenyl-4-ylcarbamate (1.30mmol) in MeOH (4 mL) was added HCl/dioxane (8 mL) at 0° C. It wasstirred at 0° C. to room temperature for 18 hours. It was concentratedin vacuo and the residue was purified by Prep-HPLC to give compound 016as a white solid (73 mg, yield: 15%, lot SP-0017146-033). LCMS:m/z=374.1 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 9.88 (s, 1H), 8.37 (d, J=3.0Hz, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.90 (d, J=2.5 Hz, 1H), 7.56 (d, J=7.4Hz, 2H), 7.45 (dd, J=8.8, 2.8 Hz, 1H), 7.40 (t, J=7.7 Hz, 2H), 7.28-7.24(m, 2H), 6.91 (d, J=8.0 Hz, 1H), 5.03 (s, 2H), 3.28 (m, 4H), 2.85 (m,4H).

Example 17—Synthesis of Compound 017

Step 1:

A mixture of lithium 5-(4-(tert-butoxycarbonyl)piperazin-1-yl)picolinate(1.30 mmol) and HATU (545 mg, 1.43 mmol) in DMF (4 mL) was stirred atroom temperature for 30 minutes. Then tert-butyl2-amino-4-(thiophen-2-yl)phenylcarbamate (416 mg, 1.43 mmol) was added.It was stirred at room temperature for 2 hours. Then it was poured intowater. The resulting solid was collected by filtration and dried invacuo to give the crude compound 5. LCMS: m/z=580.2 (M+H)⁺.

Step 2:

To a mixture of tert-butyl3-(5-(4-methylpiperazin-1-yl)picolinamido)biphenyl-4-ylcarbamate (1.30mmol) in MeOH (4 mL) was added HCl/dioxane (8 mL) at 0° C. It wasstirred at 0° C. to room temperature for 18 hours. It was concentratedin vacuo and the residue was purified by Prep-HPLC to give compound 017as a white solid (75 mg, yield: 15%, lot SP-0017146-032). LCMS:m/z=380.1 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 9.86 (s, 1H), 8.37 (d, J=3.0Hz, 1H), 7.95 (d, J=8.5 Hz, 1H), 7.88 (d, J=2.0 Hz, 1H), 7.45 (dd,J=8.5, 3.0 Hz, 1H), 7.37 (d, J=5.5 Hz, 1H), 7.25 (m, 2H), 7.06 (dd,J=5.0, 4.0 Hz, 1H), 6.85 (d, J=8.5 Hz, 1H), 5.08 (s, 2H), 3.28 (m, 4H),2.85 (m, 4H).

Example 18—Synthesis of Compound 018

Step 1:

A mixture of lithium 5-(4-(tert-butoxycarbonyl)piperazin-1-yl)picolinate(200 mg, 0.56 mmol) and HATU (214 mg, 0.56 mmol) in DMF (2 mL) wasstirred at room temperature for 15 minutes. Tert-butyl2-amino-4-(furan-2-yl)phenylcarbamate (86 mg, 0.31 mmol) was added. Itwas stirred at room temperature for 18 hours. Water was added. Theresulting solid was collected by filtration and dried in vacuo to givecrude compound 3 as an off-white solid (170 mg, 97%).

Step 2:

At 0° C., to a solution of tert-butyl4-(6-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate (170 mg, 0.30mmol) in DCM (2 mL) was added TFA (1 mL). It was stirred at 0° C. for 2h and then concentrated in vacuo. The residue was purified by Prep-HPLCto obtain compound 018 as a yellow solid (53 mg, 47%, two steps, lotSP-0017146-122). LCMS: m/z=364.1 (M+H)+. 1H NMR (400 MHz, DMSO) δ 9.83(s, 1H), 8.37 (d, J=2.8 Hz, 1H), 7.95 (d, J=8.8 Hz, 1H), 7.91 (d, J=1.2Hz, 1H), 7.62 (d, J=1.6 Hz, 1H), 7.45 (dd, J=8.8, 2.8 Hz, 1H), 7.29 (dd,J=8.4, 2.0 Hz, 1H), 6.85 (d, J=8.4 Hz, 1H), 6.61 (d, J=3.2 Hz, 1H), 6.52(m, 1H), 5.09 (s, 2H), 3.28 (m, 4H), 2.85 (m, 4H).

Example 19—Synthesis of Compound 019

Step 1:

A mixture of lithium 5-(4-(tert-butoxycarbonyl)piperazin-1-yl)picolinate(200 mg, 0.56 mmol) and HATU (214 mg, 0.56 mmol) in DMF (3 mL) wasstirred at room temperature for 15 minutes. Tert-butyl2-amino-4-(furan-3-yl)phenylcarbamate (86 mg, 0.31 mmol) was added. Itwas stirred at room temperature for 18 hours. Water was added. Theresulting solid was collected by filtration and dried in vacuo to givecrude compound 3 as an off-white solid (170 mg, 97%).

Step 2:

At 0° C., to a solution of tert-butyl4-(6-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl)phenylcarbamoyl)pyridin-3-yl)piperazine-1-carboxylate (170 mg, 0.30mmol) in DCM (1.5 mL) was added TFA (1 mL). It was stirred at 0° C. for2 hours. Then it was concentrated in vacuo. The residue was purified byprep-HPLC to obtain compound 019 as a yellow solid (67 mg, yield: 59%,two steps, lot SP-0017146-120). LCMS: m/z=364.1 (M+H)+. 1H NMR (400 MHz,DMSO) δ 9.83 (s, 1H), 8.36 (d, J=2.8 Hz, 1H), 7.96 (s, 1H), 7.95 (d,J=8.8 Hz, 1H), 7.74 (d, J=2.0 Hz, 1H), 7.67 (m, 1H), 7.44 (dd, J=8.8,2.8 Hz, 1H), 7.19 (dd, J=8.0, 2.0 Hz, 1H), 6.83 (d, J=8.0 Hz, 1H), 6.80(d, J=0.8 Hz, 1H), 4.93 (s, 2H), 3.28 (m, 4H), 2.85 (m, 4H).

Example 20—Synthesis of Compound 020

Step 1:

A mixture of compound 4 (224 mg, 0.63 mmol) and HATU (240 mg, 0.63 mmol)in DMF (5 mL) was stirred at room temperature for 15 minutes. Thencompound 5 (100 mg, 0.35 mmol) was added. It was stirred at roomtemperature for 18 hours. Then it was poured into water. The mixture wascollected by filtration and dried in vacuo to give the compound 6. LCMS:m/z=575.3 (M+H)⁺.

Step 2:

To a mixture of compound 6 (0.36 mmol) in MeOH (1 mL) was addedHCl/dioxane (2.5 mL) at 0° C. It was stirred at 0° C. to roomtemperature for 18 hours. Then it was concentrated in vacuo and theresidue was purified by prep-HPLC to give compound 020 as a light yellowsolid (79 mg, free amine, yield: 33%, two steps, lot SP-0017146-109).LCMS: m/z=375.2 (M+H)⁺. 1H NMR (400 MHz, DMSO) δ 9.87 (s, 1H), 8.52 (m,2H), 8.37 (d, J=2.8 Hz, 1H), 7.99 (d, J=2.4 Hz, 1H), 7.95 (d, J=8.8 Hz,1H), 7.58 (m, 2H), 7.45 (m, 2H), 6.92 (d, J=8.0 Hz, 1H), 5.30 (s, 2H),3.28 (m, 4H), 2.85 (m, 4H).

Example 21—Synthesis of Compound 021

Step 1:

To 2-amino-3-(4-bromophenyl)propanoic acid (50 g, 0.2 mol) in ethanol (1L) was added dropwise SOCl₂ (30 ml) at 0° C. After addition wascompleted, the mixture was refluxed for overnight. The reaction wasstopped, the mixture was evaporated to dryness, to the residue was addedEA (500 ml) and saturated aqueous NaHCO₃ (500 ml). The organic layer wasseparated out, the water was extracted with EA (200 ml), the combinedorganic layer was washed with brine and dried with Na₂SO₄, evaporated todryness and the residue was used in next step without furtherpurification. 45 g of compound 2 as a yellow solid was obtained. Yield:81%. LCMS: 99% UV-214, [M+H]:272

Step 2:

To a flask containing ethyl 2-amino-3-(4-bromophenyl)propanoate (45 g,166 mmol) in CH₃CN (500 ml) was added HCOONH₄ (95 g, 1.5 mol) at roomtemperature, followed by stirring at 90° C. overnight. After the solventwas evaporated, to the residue was added water (1 L) and then extractedby EA (300 ml×3). The combined organic layer was washed with brine,dried by anhydrous Na₂SO₄, and concentrated in vacuo. The resultingresidue was purified by silica gel chromatography (Elution:FromPE/EA=4:1 to 3:1) to afford ethyl3-(4-bromophenyl)-2-formamidopropanoate as a red solid (39 g, yield:80%). LCMS: 92% UV-214, [M+H]: 300

Step 3:

To a mixture of ethyl-3-(4-bromophenyl)-2-formamidopropanoate (39 g, 130mmol) in DCM (500 mL) was added (COCl)₂ (18 g, 143 mmol). The reactionmixture was stirred at room temperature for 30 minutes. Then thereaction solution was cooled to 0° C. and FeCl₃ (26 g, 163 mmol) wasadded into the solution, followed by stirring at room temperatureovernight. After the solution was extracted by CH₂Cl₂, the combinedorganic layers were concentrated in vacuo to afford a black oil. The oilwas dissolved in the EtOH (300 ml), and concentrated H₂SO₄ was addedinto the solution and refluxed at 80° C. overnight. The solution waspoured into saturated NaHCO₃ and extracted by EA. After drying withanhydrous Na₂SO₄, the solution was concentrated in vacuo, and theresidue was purified by silica gel chromatography (PE/EA=8:1 to 5:1) toafford ethyl 7-bromoisoquinoline-3-carboxylate as a yellow solid (5.2 g,yield: 15 percent). LCMS: 95% UV-214, [M+H]: 280

Step 4:

A mixture of compound 4 (5.2 g, 18.6 mmol), tert-butylpiperazine-1-carboxylate (4.2 g, 22.4 mmol), Pd₂(dba)₃ (920 mg, 1 mmol),RuPhos (950 mg, 2 mmol) and Cs₂CO₃ (12 g, 37 mmol) in dioxane (150 mL)was stirred at 100° C. under N₂ atmosphere for 4 hours. The mixture wascooled, filtered, and concentrated to obtain a residue, which waspurified by combiflash (Elution, PE:EA=40%) to afford compound 5 (4.5 g,63%) as a yellow solid. LCMS: 98% UV-214, [M+H]: 386

Step 5:

A solution of compound 5 (4.5 g, 11.7 mmol) in MeOH (50 mL) and THF (50ml) was added the aqueous of NaOH (2.4 g, 58.5 mmol) in water 30 ml andstirred at 60° C. for 3 hours. The mixture was concentrated to obtain aresidue, to the residue was added water (100 mL), and then adjusted pHto about 6 using HCl (0.5N) carefully, yellow solid was separated out,the mixture was filtered, the solid was washed by water (50 ml) anddried to give compound 6 as a yellow solid (3.5 g, yield: 83%). LCMS:98.9% UV-214, [M+H]: 358

Step 6:

A mixture of compound 6 (357 mg, 1 mmol), benzene-1,2-diamine (100 mg,0.9 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (380 mg, 2 mmol) in pyridine (10 mL) was stirred at roomtemperature for overnight. After completed, the mixture wasconcentrated, poured into water (100 mL), filtered to obtain compound 7(328 mg, crude) as a yellow solid.

Step 7:

A mixture of compound 7 (328 mg, crude) and TFA (3 mL) in DCM (3 mL) wasstirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 021 (101 mg, 29%, 2 steps) as a yellowsolid. LCMS: m/z=348 (M+H)⁺. ¹HNMR (500 MHz, DMSO): 3.501 (t, 4H), 3.88(t, 4H), 7.45 (t, 1H), 7.53 (t, 2H), 7.61 (d, 1H), 8.146 (dd, 1H), 8.34(d, 1H), 9.17 (s, 1H), 9.57 (s, 1H).

Example 22—Synthesis of Compound 022

Step 1:

A mixture of compound 1 (1.5 g, 4.2 mmol), tert-butyl3-aminobiphenyl-4-ylcarbamate (1.43 g, 5 mmol), HATU (2.4 g, 6.3 mmol),DIPEA (1.1 g, 8.4 mmol) in DCM (100 ml) was stirred at room temperaturefor 1 hour. The mixture was evaporated to be dry, the residue waspurified by silica gel column chromatography (DCM:MeOH=20:1) to givecompound 2 as a yellow solid (1.6 g, Yield: 62%). LCMS: 97% UV-214,[M+H]: 624

Step 2:

To a solution of compound 2 (1.6 g, 2.57 mmol) in DCM (100 ml) was added4N HCl in dioxane (30 ml), the mixture was stirred at room temperaturefor 2 h, the reaction was stopped, the mixture was filtered, the solidwas washed by ether (20 ml) and dried to give compound 022 (1.29 g, HClsalt, yield: 83%) as a yellow solid. LCMS: 100% UV-214, [M+H]: 424.¹HNMR (500 MHz, MeOD): 3.506 (t, 4H), 3.881 (t, 4H), 7.453 (t, 1H),7.527 (t, 2H), 7.671 (d, 1H), 7.737 (d, 2H), 7.811 (dd, 1H), 7.924 (m,2H), 8.146 (dd, 1H), 8.343 (d, 1H), 9.171 (s, 1H), 9.574 (s, 1H).

Example 23—Synthesis of Compound 023

Step 1:

To a solution of compound 1 (20 g, 0.087 mol) and ethyl3,3-diethoxypropionate (33 g, 0.17 mol) in EtOH (500 mL) was addedSnCl₂-2H2O (87 g, 0.39 mol), the mixture was stirred at 90° C. for 12hours. TLC monitored completion. After cooled, evaporated to obtain thecrude product. Then put into EA (600 mL), H₂O (300 mL), Na₂CO₃ (32 g)and stirred for 1 hour. The organic layer was separated, dried, filteredand concentrated to obtain compound 2 (15 g, 92%) as a yellow solid.

Step 2:

A mixture of compound 2 (560 mg, 2 mmol), N-boc-piperazine (1.3 g, 7mmol), Pd₂(dba)₃ (200 mg, 0.2 mmol), Ruphos (200 mg, 0.4 mmol), Cs₂CO₃(1.4 g, 7.3 mmol) in toluene (20 mL) using microwave (80° C. 10 min,110° C. 30 min, 145° C. 60 min). TLC monitored completion. The mixturewas added EA (30 mL), and H₂O (30 mL). Filtered off through diatomite.The organic layer was separated, washed, dried, filtered andconcentrated to obtain compound 3. Washed with PE to obtain purifiedcompound 3 (620 mg, ˜100%) as a yellow solid.

Step 3:

A mixture of compound 3 (600 mg, 1.56 mol) and 2M NaOH (5 ml) in MeOH(10 ml) and THF (10 ml) was stirred 70° C. for 4 hours. TLC monitoredcompletion. The mixture concentrated to a residue, added water (30 mL)and citric acid to adjust to PH=4-5, extracted with EA. The organiclayer was separated, washed, dried, filtered and concentrated to obtaincompound 4 (500 mg, 89.2%).

Step 4:

To a solution of compound 4 (107 mg, 0.30 mmol), tert-butyl(2-amino-4-(thiophen-2-yl)phenyl)carbamate (70 mg, 0.3 mmol) in 10 mlDMF was added HOAT (81.6 mg, 0.6 mmol), EDCI (114.6 mg, 0.6 mmol), DIPEA(193.5 mg, 1.5 mmol). The mixture was stirred at 60° C. for overnight.LCMS monitored completion, added water (10 ml) extracted with EA. Theorganic layer was separated, washed, dried, filtered and concentrated toobtain crude product. Purified by Prep-TLC to obtain compound 5 (95 mg,56%)

Step 5:

A mixture of compound 5 (95 mg, 0.16 mmol) and 2 ml TFA in 5 ml DCM wasstirred room temperature for 2 hours, LCMS monitored completion.Evaporated to get the crude product and purified by Prep-HPLC to obtaincompound 023 (20 mg, 34.2%). LCMS: m/z=430 (M+H)⁺. ¹H NMR (400 MHz,MeOD) δ 9.29 (s, 1H), 8.86 (s, 1H), 7.99 (d, J=9.2 Hz, 1H), 7.62-7.59(m, 2H), 7.47-7.44 (m, 2H), 7.40 (s, 1H), 7.28 (d, J=8.4 Hz, 1H), 7.19(d, J=3.6 Hz, 1H), 7.09 (m, 1H), 3.72 (m, 4H), 3.45 (m, 4H)

Example 24—Synthesis of Compound 024

Step 1:

A mixture of7-(4-(tert-butoxycarbonyl)piperazin-1-yl)quinoline-3-carboxylic acid(139 mg, 0.39 mmol) and EDCI (151 mg, 0.79 mmol) in pyridine (6 mL) wasstirred at room temperature for 20 minutes. Then tert-butyl2-amino-4-(furan-2-yl)phenylcarbamate (107 mg, 0.39 mmol) was added. Itwas stirred at room temperature for 16 hours. Then it was concentratedin vacuo. The residue was purified by prep-TLC to give compound 3 (143mg, 48%) as an oil. LCMS: m/z=614.3 (M+H)+.

Step 2:

To a mixture of tert-butyl4-(3-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenyl-carbamoyl)quinolin-7-yl)piperazine-1-carboxylate(143 mg, 0.23 mmol) in DCM (1 mL) was added TFA (1 mL) at 0° C. It wasstirred at 0° C. to room temperature for 18 hours. It was concentratedin vacuo and the residue was purified by prep-HPLC to give compound 024as a grey solid (37 mg, TFA salt, lot SP-0017146-056). LCMS: m/z=414.2(M+H)⁺. ¹H NMR (400 MHz, MeOD) δ 9.29 (s, 1H), 8.86 (s, 1H), 7.99 (d,J=9.2 Hz, 1H), 7.62-7.59 (m, 2H), 7.47-7.44 (m, 2H), 7.40 (s, 1H), 6.96(d, J=8.4 Hz, 1H), 6.56 (d, J=3.6 Hz, 1H), 6.47 (m, 1H), 3.72 (m, 4H),3.45 (m, 4H).

Example 25—Synthesis of Compound 025

Step 1:

A mixture of compound 3 (178 mg, 0.5 mmol), tert-butyl(2-amino-4-(furan-3-yl)phenyl)carbamate (123 mg, 0.45 mmol), and EDCI(573 mg, 3 mmol) in pyridine (5 mL) was stirred at room temperature forovernight and monitored by TLC. After pouring into water and extractingwith EA (2×100 mL), the organic layer was separated, dried, and purifiedby prep-TLC (PE/EA 1/1) to afford compound 4 (140 mg, 51%) as anoff-white solid.

Step 2:

To a solution of compound 4 (140 mg, 0.23 mmol) in DCM (5 mL) was addedTFA (2 mL), the mixture was stirred at room temperature for 2 hours.After completed, the resulting mixture was concentrated and alkalifiedwith (aqueous) NaHCO₃ to pH=8. The precipitate was collected and washedby water to afford compound 025 as an off-white solid (19 mg, 14%).LCMS: m/z=364.2 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 10.10 (s, 1H), 9.34 (s,1H), 8.96 (s, 1H), 8.89 (s, 2H), 8.06 (d, J=9.2 Hz, 1H), 8.01 (s, 1H),7.69 (t, J=1.6 Hz, 1H), 7.65 (dd, J=9.3, 2.2 Hz, 1H), 7.46 (s, 1H), 7.40(d, J=1.9 Hz, 1H), 7.33 (dd, J=8.3, 2.0 Hz, 1H), 6.90 (d, J=8.3 Hz, 1H),6.84 (s, 1H), 3.70-3.66 (m, 4H), 3.32 (s, 4H).

Example 26—Synthesis of Compound 026

Step 1:

A mixture of compound 3 (357 mg, 1 mmol), tert-butyl(2-amino-4-(pyridin-4-yl)phenyl)carbamate (260 mg, 0.9 mmol), and EDCI(381 mg, 2 mmol) in pyridine (5 mL) was stirred at room temperature forovernight and monitored by TLC. Poured into water and extracted with EA(2×100 mL), the organic layer was separated, dried, and purified byprep-TLC (PE/EA 1/1) to afford compound 4 (240 mg, 39%) as an off-whitesolid.

Step 2:

To a solution of compound 4 (240 mg, 0.23 mmol) in DCM (5 mL) was addedTFA (2 mL), the mixture was stirred at room temperature for 2 hours.After completion, the resulting mixture was concentrated and madealkaline with (aqueous) NaHCO₃ to pH=8. The precipitate was collectedand washed by water to afford compound 026 as an off-white solid (100mg, 62%). LCMS: m/z=425 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 9.909 (s, 1H),9.28 (s, 1H), 8.79 (s, 1H), 8.52 (s, 1H), 7.92-7.90 (d, J=8 Hz, 1H),7.74 (s, 1H), 7.61-7.53 (m, 4H), 7.26 (s, 1H), 6.92-6.90 (d, J=8 Hz,1H), 5.48 (s, 2H), 2.90 (m, 4H), 2.51 (s, 4H).

Example 27—Synthesis of Compound 027

Step 1:

6-(4-methylpiperazin-1-yl)nicotinic acid (132 mg, 0.6 mmol), tert-butyl2-amino-4-(pyridin-4-yl) phenylcarbamate (172 mg, 0.6 mmol) and EDCI(346 mg, 1.8 mmol) were added into pyridine (5 ml). The mixture wasstirred for overnight at room temperature. When the reaction finished,it was extracted by EA and washed by citric acid, NaHCO₃ and saturatedbrine. Then the organic layer was concentrated to afford compound 2 (300mg, crude).

Step 2:

Tert-butyl 2-(6-(4-methylpiperazin-1-yl) nicotinamido)-4-(pyridin-4-yl)phenylcarbamate (300 mg, crude) was added into DCM (2 ml). Then TFA (2ml) was added. The mixture was stirred at room temperature for 1 hours.It was purified by Prep-HPLC (base method). Compound 027 was obtained asa yellow solid (40 mg, 13% yield, lot SP-0017467-055). LCMS: m/z=389.2(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.76 (d, J=2.3 Hz, 1H),8.53-8.45 (m, 2H), 8.10 (dd, J=9.0, 2.5 Hz, 1H), 7.67 (d, J=2.1 Hz, 1H),7.58 (d, J=6.2 Hz, 2H), 7.50 (dd, J=8.4, 2.2 Hz, 1H), 6.90 (dd, J=13.9,8.7 Hz, 2H), 5.36 (s, 2H), 3.67-3.58 (m, 4H), 2.43-2.35 (m, 4H), 2.22(s, 3H).

Example 28—Synthesis of Compound 028

Step 1:

Under N₂, a mixture 1-methylpiperazine (232 mg, 2.31 mmol) and methyl5-bromopicolinate (500 mg, 2.31 mmol),tris(dibenzylideneacetone)dipalladium (53 mg, 0.058 mmol), Ru-Phos (108mg, 0.23 mmol) and cesium carbonate (2.26 g, 6.94 mmol) in toluene (20mL) was heated at 100° C. for 18 hours. Then it was cooled to roomtemperature and filtrated. The filtrate was concentrated in vacuo. Andthe residue was washed with PE-EA to give compound 3 as a white solid(528 mg, 100%).

Step 2:

At 0° C., to a mixture of methyl 5-(4-methylpiperazin-1-yl)picolinate(528 mg, 2.24 mmol) and LiOH (188 mg, 4.49 mmol) in THF (6 mL), MeOH (1mL) and water (3 mL). It was stirred at 0° C. to room temperature for 18hours. Then it was concentrated in vacuo to give crude compound 4 as aLi salt (294 mg). LCMS: m/z=222.1 (M+H)⁺.

Step 3:

A mixture of compound 4 (143 mg, 0.63 mmol) and HATU (240 mg, 0.63 mmol)in DMF (10 mL) was stirred at room temperature for 10 minutes. Thencompound 5 (100 mg, 0.35 mmol) was added. It was stirred at roomtemperature for 18 hours. Then it was poured into water. The resultingsolid was collected by filtration and dried in vacuo to give the crudecompound 6 (150 mg). LCMS: m/z=489.3 (M+H)+.

Step 4:

To a mixture of compound 6 (0.36 mmol) in MeOH (1 mL) was addedHCl/dioxane (4 M, 2 mL) at 0° C. It was stirred at 0° C. to roomtemperature for 18 hours. Then it was concentrated in vacuo and theresidue was purified by Prep-HPLC to give compound 028 as a yellow solid(59 mg, free amine, yield: 24%, two steps, lot SP-0017146-108). LCMS:m/z=389.3 (M+H)⁺. 1H NMR (400 MHz, DMSO) δ 9.88 (s, 1H), 8.52 (m, 2H),8.39 (d, J=2.4 Hz, 1H), 7.99 (d, J=2.0 Hz, 1H), 7.96 (d, J=9.2 Hz, 1H),7.58 (m, 2H), 7.47 (m, 2H), 6.92 (d, J=8.0 Hz, 1H), 5.30 (s, 2H), 3.38(m, 4H), 2.47 (m, 4H).

Example 29—Synthesis of Compound 029

Step 1:

A mixture of compound 1 (100 mg, 0.37 mmol), tert-butyl2-amino-4-(furan-2-yl)phenyl-carbamate (151 mg, 1.5 eq.) and EDCI (550mg, 7.0 eq.) in pyridine (5 ml) were stirred at 25° C. overnight. It wasconcentrated and extracted with EA (2×50 ml), washed by brine (2×60 ml).It was purified by prep-TLC (PE:EA=1:3). Compound 2 (150 mg, Yield: 39%)was obtained as a yellow oil.

Step 2:

To a solution of compound 2 (150 mg, 0.28 mmol) in DCM (2 ml) was addedTFA (0.5 ml) at room temperature. The reaction was stirred for 2 hours.Then it was concentrated. Compound 029 was obtained as a yellow solid(33 mg, 27%). LCMS: m/z=428 (M+H)⁺ ¹H NMR (400 MHz, MeOD) δ 9.17 (s,1H), 8.50 (s, 1H), 7.99 (d, J=9.1 Hz, 1H), 7.90 (d, J=1.8 Hz, 1H),7.75-7.68 (m, 1H), 7.49 (s, 2H), 7.43 (dd, J=8.3, 1.9 Hz, 1H), 6.97 (d,J=8.4 Hz, 1H), 6.59 (d, J=3.3 Hz, 1H), 6.48 (dd, J=3.2, 1.8 Hz, 1H),3.62 (s, 4H), 3.19 (s, 4H), 2.78 (s, 3H).

Example 30—Synthesis of Compound 030

Step 1:

To a solution of compound 1 (432 mg, 4.6 mmol) and ethyl2-bromothiazole-5-carboxylate (1 g, 4.2 mmol), Et₃N (1.27 g, 12.6 mmol)in DMF (20 ml) was stirred at 90° C. for overnight. The solvent wasevaporated off and extracted by EA (20 ml×2), washed with water, brine,concentrated in vacuo to afford compound 2 as a yellow solid (1.2 g,crude).

Step 2:

To a solution of compound 2 (1.2 g, 4.7 mmol) NaOH (2.5 ml, 2M) in EtOH(10 ml) was stirred at 60° C. for 2 hours. The solvent was evaporatedoff, adjusted to pH<7, concentrated to afford compound 3 (2.0 g,containing some NaCl).

Step 3:

To a solution of compound 3 (454 mg, 2 mmol) tert-butyl(2-amino-4-(thiophen-2-yl)phenyl)carbamate (290 mg, 1 mmol) EDCI (382mg, 2 mmol) in pyridine (10 ml). The mixture was stirred at roomtemperature for overnight. The solvent was evaporated off, extracted byEA (15 ml×2) and washed with water to afford compound 4 as a white solid(210 mg, crude).

Step 4:

To a solution of compound 4 (210 mg, 0.42 mmol) in DCM (5 ml) was addedTFA (2 ml). The mixture was stirred room temperature for 2 hours. Thesolvent was evaporated off, adjusted to pH<7 and filtered to affordcompound 030 as a white solid (58 mg, 34%). LCMS: m/z=500 (M+H)⁺ ¹H NMR(500 MHz, DMSO) δ 9.53 (s, 1H), 8.04 (s, 1H), 7.32-7.23 (m, 4H), 6.80(s, 1H), 6.79-6.76 (d, J=8 Hz, 1H), 5.13 (s, 2H), 3.505-3.485 (s, 4H),2.42-2.41 (s, 4H), 2.227 (s, 3H).

Example 31—Synthesis of Compound 031

Step 1:

A mixture of methyl 2-bromo-4-methylthiazole-5-carboxylate (472 mg, 2mmol), tert-butyl piperazine-1-carboxylate (1.12 g, 6 mmol), DIPEA (1.29g, 10 mmol) in 1,4-dioxane (30 mL) was stirred at 95° C. under N₂atmosphere for overnight. The mixture was cool and added EA (100 mL) andwater (100 mL), stirred for 30 min, the organic layer was separated,dried, concentrated to get a residue, which was washed by PE (100 mL) toobtain compound 2 (477 g, 70%) as a light yellow solid.

Step 2:

A mixture of compound 2 (341 mg, 1 mmol) and 2M NaOH (5 mL) in THF (5ml) and EtOH (5 mL) was stirred at 60° C. for 3 h The mixture wasconcentrated to get a residue, HCl (2 M) was added to adjust pH 7,filtered to give compound 3 as a white solid (298 mg, 91%).

Step 3:

A mixture of compound 3 (307 mg, 1 mmol), tert-butyl2-amino-4-(thiophen-2-yl)phenyl-carbamate (290 mg, 1 mmol), HOAT (136mg, 1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(573 mg, 3 mmol) and DMAP (244 mg, 2 mmol) in DMF (5 mL) was stirred at55° C. for overnight. The mixture was poured into water (100 mL),filtered to get a residue which was purified by Prep-TLC to obtaincompound 4 (90 mg, 15%) as a yellow solid.

Step 4:

A mixture of compound 4 (90 mg, 0.15 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 hour. The mixture was purified byPrep-HPLC to obtain compound 031 (34 mg, 57%) as a yellow solid. LCMS:m/z=400 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 8.95 (s, 1H), 8.21 (s, 1H),7.45 (s, 1H), 7.37 (m, 1H), 7.27 (m, 2H), 7.06 (m, 1H), 6.80 (m, 1H),5.06 (s, 2H), 3.43 (s, 4H), 2.85 (s, 4H).

Example 32—Synthesis of Compound 032

Step 1:

To a solution of compound 1 (860 mg, 4.6 mmol) and ethyl2-bromothiazole-5-carboxylate (1 g, 4.2 mmol), Et₃N (1.27 g, 12.6 mmol)in DMF (20 ml) was stirred at 90° C. for overnight. The solvent wasevaporated off and extracted by EA (20 ml×2), washed with water, brine,concentrated in vacuo to afford compound 2 as a yellow solid (1.8 g,crude).

Step 2:

To a solution of compound 2 (500 mg, 1.46 mmol) NaOH (2 ml, 2M) in EtOH(10 ml) was stirred at 60° C. for 2 hours. The solvent was evaporatedoff, adjusted to pH<7, filtered and washed with water to afford compound3 as a white solid (394 mg, 86.1%).

Step 3:

To a solution of compound 3 (95 mg, 0.3 mmol) tert-butyl(2-amino-4-(pyridin-4-yl)phenyl)carbamate (86 mg, 0.3 mmol) EDCI (172mg, 0.9 mmol) in pyridine (4 ml). The mixture was stirred at roomtemperature for overnight. The solvent was evaporated off, extracted byEA (15 ml×2) and washed with water to afford compound 4 as a white solid(220 mg, crude).

Step 4:

To a solution of compound 4 (200 mg, 0.37 mmol) in DCM (2 ml) was addedTFA (1 ml). The mixture was stirred room temperature for 2 hours. Thesolvent was evaporated off, adjusted to pH<7 and filtered to affordcompound 032 as a white solid (68 mg, 34%). ¹H NMR (400 MHz, DMSO) δ9.54 (s, 1H), 8.50 (d, J=6.0 Hz, 2H), 8.06 (s, 1H), 7.62 (s, 1H), 7.58(d, J=6.1 Hz, 2H), 7.49 (d, J=8.3 Hz, 1H), 6.87 (d, J=8.4 Hz, 1H), 5.36(s, 2H), 3.45-3.38 (m, 4H), 2.79 (s, 4H). LCMS: m/z=382 (M+H)⁺

Example 33—Synthesis of Compound 033

Step 1:

Methyl 5-bromothiophene-2-carboxylate (663 mg, 3 mmol), tert-butylpiperazine-1-carboxylate (1.674 g, 9 mmol), Pd₂(dba)₃ (275 mg, 0.3mmol), RuPhos (140 mg, 0.3 mg) and Cs₂CO₃ (2.934 g, 9 mmol) were addedinto toluene (15 ml). The mixture was stirred at 95° C. for overnight.It was purified by column chromatography. Compound 2 was obtained as agreen solid (660 mg, 56% yield).

Step 2:

Tert-butyl 4-(5-(methoxycarbonyl) thiophen-2-yl)piperazine-1-carboxylate (640 mg, 1.96 mmol) and NaOH (2 mol/L, 4 ml)were added into EtOH (4 ml) and THF (4 ml). The mixture was stirred at60° C. for 3 hours. The pH was adjusted to 6-7 and it was filtered andwashed by Et₂O. Compound 3 was obtained as a green solid (600 mg, 87%yield).

Step 3:

5-(4-(tert-butoxycarbonyl) piperazin-1-yl) thiophene-2-carboxylic acid(94 mg, 0.3 mmol), tert-butyl 2-amino-4-(thiophen-2-yl) phenylcarbamate(87 mg, 0.3 mmol) and EDCI (173 mg, 0.9 mmol) were added into pyridine(3 ml). The mixture was stirred at room temperature for overnight. Acrude compound 4 (200 mg) was used for next step.

Step 4:

Tert-butyl 4-(5-(2-(tert-butoxycarbonylamino)-5-(thiophen-2-yl)phenylcarbamoyl) thiophen-2-yl) piperazine-1-carboxylate (180 mg, 0.32mmol) was added into DCM (2 ml). Then TFA (2 ml) was added. The mixturewas stirred at room temperature for 2 hours. When the reaction finished,the pH was adjusted to 8-9. It was extracted by EA and washed by brine,HCl. Compound 033 was afforded as HCl salt (49 mg, 42% yield, lotSP-0017467-087). LCMS: m/z=385.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 10.23(s, 1H), 9.40 (s, 2H), 8.04 (d, J=3.8 Hz, 1H), 7.72 (s, 1H), 7.51 (dd,J=22.9, 10.9 Hz, 3H), 7.29 (d, J=8.2 Hz, 1H), 7.13 (s, 1H), 6.38 (d,J=3.9 Hz, 1H). 1H NMR (400 MHz, D20) δ 7.55 (dd, J=11.5, 6.6 Hz, 2H),7.45 (s, 1H), 7.39-7.31 (m, 2H), 7.28 (d, J=8.2 Hz, 1H), 7.04 (s, 1H),6.24 (d, J=4.4 Hz, 1H), 3.46 (d, J=5.3 Hz, 4H), 3.35-3.20 (m, 4H).

Example 34—Synthesis of Compound 034

Step 1:

To a solution of compound 1 (130 mg, 0.41 mmol) and tert-butyl(2-amino-4-(furan-2-yl)phenyl)carbamate (118 mg, 0.41 mmol) in pyridine(2 ml) was added EDCI (237 mg, 1.23 mmol). The mixture was stirred atroom temperature for overnight. After completed, the solvent wasevaporated off and extracted by EA (20 ml×2), washed with water, brine,purified by prep-TLC (PE/EA=1/1) to afford compound 2 as a white solid(135 mg, 56%).

Step 2:

To a solution of compound 2 (120 mg, 0.2 mmol) in DCM (2 ml) was addedTFA (1 ml). The mixture was stirred at room temperature for 2 hours. Thesolvent was evaporated off, adjusted to pH 7-8 and purified by prep-HPLCto afford compound 034 as a white solid (28 mg, 31.8%). ¹H NMR (400 MHz,DMSO) δ 9.53 (s, 1H), 8.30 (s, 1H), 8.05 (s, 1H), 7.60 (s, 1H), 7.45 (d,J=1.7 Hz, 1H), 7.31 (dd, J=8.3, 1.8 Hz, 1H), 6.79 (d, J=8.3 Hz, 1H),6.61 (d, J=3.3 Hz, 1H), 6.52-6.48 (m, 1H), 5.14 (s, 2H), 3.43 (s, 4H),2.82 (s, 4H). LCMS: m/z=370 (M+H)⁺

Example 35—Synthesis of Compound 035

Step 1:

To a solution of tert-butyl2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenylcarbamate(1.0 g, 2.75 mmol) and 1-bromo-4-fluorobenzene (480 mg, 2.750 mmol) inDioxane (6 ml) and H₂O (3 ml) was added K₂CO₃ (560 mg, 5.50 mmol) andPd(PPh₃)₄ (160 mg, 0.14 mmol). The mixture was stirred at 100° C. underN₂ for overnight. The mixture was directly purified on a column with(PE/EA=20/1) to afford compound 2 as a yellow solid (820 mg, 90%).

Step 2:

To a solution of tert-butyl 4′-fluoro-3-nitrobiphenyl-4-ylcarbamate (800mg, 2.41 mmol) and FeCl₃ (80 mg, 0.48 mmol) in EtOH (12 ml) was added C(800 mg, 63.41 mmol) and drop wise N₂H₄H₂O (5 ml). The mixture wasstirred at 60° C. for 2 hours. The mixture was extracted with H₂O (3×100ml). The combined organic layers were washed with brine (50 ml), driedover Na₂SO₄ and concentrated in vacuo to afford compound 3 as a whitesolid (600 mg, 80%).

Step 3:

To a solution of tert-butyl 3-amino-4′-fluorobiphenyl-4-ylcarbamate (115mg, 0.38 mmol) and2-(4-(tert-butoxycarbonyl)piperazin-1-yl)quinoline-6-carboxylic acid(151 mg, 0.38 mmol) in pyridine (2.5 ml) was added EDCI (226 mg, 1.08mmol). The mixture was stirred at room temperature for overnight. Theaqueous layer was extracted with H₂O (3×100 ml). The combined organiclayers were washed with brine (50 ml), dried over Na₂SO₄ andconcentrated in vacuo to afford compound 4 as a white solid (200 mg,100%).

Step 4:

To a solution of tert-butyl4-(6-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)quinolin-2-yl)piperazine-1-carboxylate(200 mg, 0.31 mmol) in DCM (2 ml) at 0° C. was added TFA (2 ml)drop-wise. And the resulted was stirred at room temperature for 2 hours.The residue was purified by Prep-HPLC (0.1% NH₄HCO₃/water-acetonitrileto afford compound 035 as a white solid (64 mg, 47%). ¹H NMR (400 MHz,DMSO) δ 9.77 (s, 1H), 8.40 (d, J=1.9 Hz, 1H), 8.11 (dd, J=13.3, 5.6 Hz,2H), 7.62-7.55 (m, 3H), 7.53 (d, J=2.0 Hz, 1H), 7.34-7.26 (m, 2H), 7.22(t, J=8.9 Hz, 2H), 6.87 (d, J=8.4 Hz, 1H), 5.13 (s, 2H), 3.81-3.53 (m,4H), 2.78 (dd, J=33.1, 28.1 Hz, 4H). LCMS: m/z=442.3 (M+H)⁺.

Example 36—Synthesis of Compound 036

Step 1:

6-(4-(tert-butoxycarbonyl) piperazin-1-yl) nicotinic acid (144 mg, 0.47mmol), tert-butyl 3-amino-4′-fluorobiphenyl-4-ylcarbamate (100 mg, 0.33mmol), HATU (357 mg, 0.94 mmol) and DIPEA (0.25 ml, 1.41 mmol) wereadded into THF (5 ml). The mixture was stirred for 2 days at roomtemperature. When the reaction finished, it was extracted by EA andwashed by brine. It was purified by column chromatography (PE/EA=2:1). AWhite solid was afforded as compound 2 (100 mg, 51% yield).

Step 2:

Tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)pyridin-2-yl) piperazine-1-carboxylate (100 mg, 0.17 mmol) was dissolvedin DCM (0.5 ml). Then TFA (2 ml) was added. The mixture was stirred for2 hours. When the reaction finished, the pH was adjusted to 10. And itwas extracted by EA. The organic layer was concentrated and washed byEt₂O. A white solid was afforded as compound 036 (52 mg, 79% yield, lotSP-0017467-103). LCMS: m/z=392.2 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.51(s, 1H), 8.75 (s, 1H), 8.09 (s, 1H), 7.53 (d, J=36.6 Hz, 3H), 7.25 (d,J=25.7 Hz, 3H), 6.86 (s, 2H), 5.09 (s, 2H), 3.55 (s, 4H), 2.77 (s, 4H).

Example 37—Synthesis of Compound 037

Step 1:

To a solution of compound 1 (200 mg, 0.66 mmol) and tert-butyl(3-amino-4′-fluoro-[1,1′-biphenyl]-4-yl)carbamate (100 mg, 0.33 mmol)EDCI (189 mg, 0.99 mmol) in pyridine (2 ml) was stirred at roomtemperature for overnight. The solvent was evaporated off and extractedby EA (20 ml×2), then it was washed with HCl (1N, 10 ml), water andbrine, concentrated in vacuo to afford compound 2 as a yellow solid (165mg, 83.7%).

Step 2:

To a solution of compound 2 (165 mg, 0.27 mmol) in DCM (1 ml) was addedTFA (1 ml). The mixture was stirred room temperature for 2 hours. Thesolvent was evaporated off, adjusted to pH 7-8 and washed with Et₂O (10ml) to afford compound 037 as a yellow solid (101 mg, 92.6%). ¹H NMR(400 MHz, MeOD) δ 7.99 (s, 1H), 7.60-7.54 (m, 2H), 7.41 (d, J=2.1 Hz,1H), 7.34 (dd, J=8.3, 2.2 Hz, 1H), 7.12 (t, J=8.8 Hz, 2H), 6.98-6.94 (m,1H), 3.59-3.53 (m, 4H), 2.99-2.93 (m, 4H). LCMS: m/z=398 (M+H)⁺

Example 38—Synthesis of Compound 038

Step 1:

Under N₂, a mixture of tert-butyl piperazine-1-carboxylate (448 mg, 2.41mmol), methyl 5-bromopicolinate (520 mg, 2.41 mmol), Ruphos (109 mg,0.24 mmol), Pd₂(dba)₃ (55 mg, 0.06 mmol) and Cs₂CO₃ (2.35 g, 7.23 mmol)in toluene (20 ml) was heated at 100° C. for overnight. Then it wascooled to room temperature and filtered. The residue was concentrated invacuum to afford a yellow solid (750 mg, 96%).

Step 2:

A mixture of tert-butyl4-(6-(methoxycarbonyl)pyridin-3-yl)piperazine-1-carboxylate (750 mg,4.67 mmol) and LiOH (200 mg, 4.67 mmol) in MeOH (1 ml), THF (6 ml) andwater (3 ml) was stirred at 0° C. for overnight. The reaction mixturewas concentrated in vacuum. Then it was washed by EA and concentrated invacuum to afford a yellow solid (800 mg, 100%).

Step 3:

To a solution of tert-butyl2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenylcarbamate(1.5 g, 4.12 mmol) and 1-bromo-4-fluorobenzene (717 mg, 4.12 mmol) inDioxane (10 ml) and H₂O (5 ml) was added K₂CO₃ (824 mg, 8.24 mmol) andPd(pph₃)₄ (238 mg, 0.21 mmol). The mixture was stirred at 100° C. underN₂ for overnight. The mixture was directly purified on a column with(PE/EA=20/1) to afford yellow solid (1.26 g, 91%).

Step 4:

To a solution of tert-butyl 4′-fluoro-3-nitrobiphenyl-4-ylcarbamate (1.1g, 3.31 mmol) and FeCl₃ (98 mg, 0.60 mmol) in EtOH (10 ml) was addedactivate carbon (1.1 g, 87.05 mmol) and drop wise N₂H₄—H₂O (5 ml). Themixture was stirred at 60° C. for 2 h. The mixture was extracted withH₂O (3×100 ml). The combined organic layers were washed with brine (50ml), dried over Na₂SO₄ and concentrated in vacuum to afford a whitesolid (1 g, 100%).

Step 5:

To a solution of lithium5-(4-(tert-butoxycarbonyl)piperazin-1-yl)picolinate (800 mg, 2.61 mmol)and HATU (992 mg, 2.61 mmol) in DMF (10 ml) was stirred at roomtemperature for 15 minutes. Tert-butyl3-amino-4′-fluorobiphenyl-4-ylcarbamate (438 mg, 1.45 mmol) was added.The mixture was stirred at room temperature for overnight. The aqueouslayer was extracted with H₂O (3×200 ml). The combined organic layerswere washed with brine (50 ml), dried over Na₂SO₄, filtered andconcentrated in vacuum. The residue was purified by preparative TLC(silica gel, GF254 10-40 u, 25×25 cm) with petroleum ether/EtOAc (2:1)to afford a white solid (120 mg, 14%).

Step 6:

To a solution of tert-butyl4-(6-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)pyridin-3-yl) piperazine-1-carboxylate (120 mg, 0.20 mmol) in DCM (1.5ml) at 0° C. was added TFA (1.5 ml) drop wise. And the resulted wasstirred at room temperature for 2 hours. The residue mixture wasconcentrated in vacuum. The residue was purified by Prep-HPLC. The whitesolid was re-crystallized from water, dried by lyophilization to affordcompound 038 as a white solid (47 mg, 59%). LCMS: m/z=392.2 (M+H)⁺ ¹HNMR (400 MHz, DMSO) δ 9.87 (s, 1H), 8.38 (d, J=2.8 Hz, 1H), 8.20 (s,1H), 7.96 (d, J=8.8 Hz, 1H), 7.84 (d, J=2.1 Hz, 1H), 7.55 (dt, J=22.5,12.9 Hz, 2H), 7.47 (dd, J=8.9, 2.8 Hz, 1H), 7.31-7.14 (m, 3H), 6.88 (d,J=8.3 Hz, 1H), 5.02 (s, 2H), 3.41-3.35 (m, 4H), 3.02-2.95 (m, 4H).

Example 39—Synthesis of Compound 039

Step 1:

To a solution of 5-bromothiazole-2-carboxylic acid (957 mg, 4.58 mmol)in DCM (20 ml) was added oxalyl chloride (0.49 ml), containing acatalytic amount of DMF (0.11 ml) at room temperature and resultingreaction mixture was stirred at room temperature for 0.5 hours. ThenMeOH (8 ml) was added to the solution and the reaction mixture wasstirred for addition 2 hours. The reaction mixture was a saturatedaqueous solution of NaHCO₃ (50 ml) and extracted with DCM (2×50 ml), thecombined organic layer was washed with water and brine (50 ml), driedover Na₂SO₄ and concentrated in vacuum to afford a white solid (900 mg,90%).

Step 2:

Under N₂, a mixture of tert-butyl piperazine-1-carboxylate (750 mg, 4.04mmol), methyl 5-bromothiazole-2-carboxylate (900 mg, 4.04 mmol),Brettphos (215 mg, 0.40 mmol), Pd₂(dba)₃ (370 mg, 0.40 mmol) and Cs₂CO₃(3.95 g, 12.1 mmol) in toluene (20 ml) was heated at 100° C. forovernight. The residue was purified by preparative TLC (silica gel, GF254 10-40 u, 25×25 cm) with petroleum ether/EtOAc (5:6) to afford ayellow solid (210 mg, 15.9%).

Step 3:

To a solution of methyl 5-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiazole-2-carboxylate (210 mg, 0.64 mmol) and LiOH (81 mg, 1.92 mmol)in MeOH (1 ml), THF (3 ml) and H₂O (1.5 ml) was stirred at 0° C. for 5h. The reaction mixture was concentrated in vacuum to afford a yellowsolid (200 mg, 100%).

Step 4:

To a solution of lithium5-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-2-carboxylate (60 mg,0.19 mmol) and HATU (73 mg, 0.19 mmol) in DMF (1.5 ml) was stirred atroom temperature for 15 min. tert-butyl2-amino-4-(thiophen-2-yl)phenylcarbamate (31 mg, 0.11 mmol) was added.The mixture was stirred at room temperature for overnight. The residuewas purified by preparative TLC (silica gel, GF254 10-40 u, 25×25 cm)with petroleum ether/EtOAc (2:1) to afford a yellow solid (50 mg, 45%).

Step 5:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(thiophen-2-yl) phenylcarbamoyl)thiazol-5-yl) piperazine-1-carboxylate (50 mg, 0.09 mmol) in DCM (1 ml)at 0° C. was added TFA (1 ml) drop wise. The resulting mixture wasstirred at room temperature for 2 h. The reaction mixture wasconcentrated in vacuum. The pH was adjusted to around 7 by progressivelyadding NaOH (5 ml). The mixture was filtered to afford compound 039 as ayellow solid. (31 mg, 94%) LCMS: m/z=386.1 (M+H)⁺ ¹H NMR (400 MHz, DMSO)δ 9.70 (s, 1H), 7.61 (d, J=1.9 Hz, 1H), 7.37 (d, J=5.0 Hz, 1H), 7.25(dd, J=10.1, 7.1 Hz, 3H), 7.09-7.01 (m, 1H), 6.82 (d, J=8.3 Hz, 1H),5.14 (s, 2H), 3.21-3.12 (m, 4H), 2.90-2.80 (m, 4H), 1.24 (s, 1H).

Example 40—Synthesis of Compound 040

Step 1:

Tert-butyl piperazine-1-carboxylate (3.72 g, 20 mmol), methyl5-chloropyrazine-2-carboxylate (1.72 g, 10 mmol) and DIPEA (2.02 g, 20mmol) were added into 1,4-dioxane (15 ml). The mixture was stirred forovernight at 100° C. When the reaction finished, it was extracted by EAand washed by brine. It was purified by flash column chromatography (EA,100%) to afford the yellow solid as product (6 g, crude).

Step 2:

Methyl 5-(4-(tert-butoxycarbonyl) piperazin-1-yl) pyrazine-2-carboxylate(2 g, crude) was added into a solution of EtOH (5 ml), THF (5 ml) andNaOH (2 N, 5 ml). The mixture was stirred at 60° C. for 2 h. When thereaction finished, the pH was adjusted to 1 and it was extracted by EA,concentrated to afford the pink solid. (300 mg, 15% yield, 2 steps).

Step 3:

5-(4-(tert-butoxycarbonyl) piperazin-1-yl) pyrazine-2-carboxylic acid(200 mg, 0.65 mmol), tert-butyl 2-amino-4-(thiophen-2-yl)phenylcarbamate (157 mg, 0.54 mmol) and EDCI (311 mg, 1.62 mmol) wereadded into Py (5 ml). The mixture was stirred at room temperature forovernight. When the reaction finished, it was concentrated and washedwith Et2O. The yellow solid was collected as product (310 mg, 65%yield).

Step 4:

Tert-butyl 4-(5-(2-(tert-butoxycarbonylamino)-5-(thiophen-2-yl)phenylcarbamoyl) pyrazin-2-yl) piperazine-1-carboxylate (300 mg, 0.52mmol) was dissolved into DCM (2 ml). Then TFA (2 ml) was added. Themixture was stirred at room temperature for 2 h. When the reactionfinished, it was adjusted to pH=10 and extracted by EA. It wasconcentrated and washed by Et₂O to afford compound 040 as a yellow solidproduct (140 mg, 69% yield, Lots SP-0017467-141). LCMS: m/z=381.2(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.68 (s, 1H), 8.72 (s, 1H), 8.34 (s,1H), 7.79 (s, 1H), 7.37 (d, J=4.7 Hz, 1H), 7.26 (d, J=8.7 Hz, 2H), 7.06(s, 1H), 6.84 (d, J=8.1 Hz, 1H), 5.10 (s, 2H), 3.68 (s, 4H), 2.83 (s,4H).

Example 41—Synthesis of Compound 041

Step 1:

To a solution of benzyl 2-aminopropanoate (10 g, 46.52 mmol), methyl2-chloro-2-oxoacetate (6.4 g, 51.16 mmol) and TEA (14 g, 139.56 mmol) inDCM (200 ml) was stirred at room temperature for 2 h. The mixture wasdirectly purified on column chromatograph on silica gel (200-300 mesh,eluting with petroleum ether/EtOAc=4:1) to afford yellow oil (8.7 g,70.7%).

Step 2:

Under H₂, a mixture of benzyl 2-(2-methoxy-2-oxoacetamido)propanoate(8.7 g, 32.8 mmol), Pd/C (400 mg, 3.8 mmol) was stirred at roomtemperature for overnight. The residue mixture was concentrated invacuum to afford colorless oil (5.5 g, 96%).

Step 3:

To a solution of tert-butyl piperazine-1-carboxylate (6.5 g, 34.57 mmol)and 2-(2-methoxy-2-oxoacetamido)propanoic acid (5.5 g, 31.43 mmol) in Py(100 ml) was added EDCI (12 g, 62.86 mmol). The mixture was stirred atroom temperature for overnight. The mixture was directly purified oncolumn chromatograph on silica gel (200-300 mesh, eluting withpetroleum/DCM/EtOAc=10:10:1) to afford yellow solid (5.1 g, 48%).

Step 4:

Under a nitrogen atmosphere, a mixture of tert-butyl4-(2-(2-methoxy-2-oxoacetamido)propanoyl)piperazine-1-carboxylate (750mg, 2.19 mmol) and LR (750 mg, 2.62 mmol) in toluene (15 mL) was stirredat 100° C. for 3 h. The residue was purified by preparative TLC (silicagel, GF 254 10-40 u, 25×25 cm) with petroleum ether/EtOAc (1:1) toafford a yellow solid (300 mg, 40%).

Step 5:

To a solution of methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylthiazole-2-carboxylate(300 mg, 0.88 mmol) and LiOH (55 mg, 1.32 mmol) in MeOH (1 ml), THF (3ml) and H₂O (1.5 ml) was stirred at 0° C. for 5 h. The reaction mixturewas concentrated in vacuum to afford a yellow solid (290 mg, 100%).

Step 6:

To a solution of5-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylthiazole-2-carboxylicacid (200 mg, 0.62 mmol) and HATU (233 mg, 0.62 mmol) in DMF (5 ml) wasstirred at room temperature for 15 min. Tert-butyl2-amino-4-(thiophen-2-yl)phenylcarbamate (99 mg, 0.35 mmol) was added.The mixture was stirred at room temperature for overnight. The residuewas purified by preparative TLC (silica gel, GF254 10-40 u, 25×25 cm)with petroleum ether/EtOAc (3:1) to afford a yellow solid (183 mg, 87%).

Step 7:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(thiophen-2-yl)phenylcarbamoyl)-4-methylthiazol-5-yl)piperazine-1-carboxylate(183 mg, 0.31 mmol) in DCM (1.5 ml) at 0° C. was added TFA (1.5 ml)dropwise. And the resulted was stirred at room temperature for 2 h. Thereaction mixture was concentrated in vacuum. The pH was adjusted toaround 7 by progressively adding NaHCO₃ (˜5 ml). The mixture wasfiltered and the filtrate was concentrated to afford a yellow solidcompound 041 (114 mg, 92%). LCMS: m/z=400.2 (M+H)⁺. ¹H NMR (400 MHz,DMSO) δ 9.89 (s, 1H), 7.55 (d, J=1.8 Hz, 1H), 7.37 (d, J=5.0 Hz, 1H),7.29 (dd, J=8.3, 1.9 Hz, 1H), 7.23 (d, J=3.3 Hz, 1H), 7.10-7.01 (m, 1H),6.82 (d, J=8.4 Hz, 1H), 5.16 (s, 2H), 3.19 (d, J=4.6 Hz, 4H), 3.06 (s,4H), 2.37 (s, 3H).

Example 42—Synthesis of Compound 042

Step 1:

To a solution of lithium5-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylthiazole-2-carboxylate(100 mg, 0.31 mmol) and HATU (116 mg, 0.31 mmol) in DMF (4 ml) wasstirred at room temperature for 15 min. Tert-butyl2-amino-4-(furan-2-yl)phenylcarbamate (47 mg, 0.17 mmol) was added. Themixture was stirred at room temperature for overnight. The residue wasadded water (˜5 mL) and the precipitate was afforded as a yellow solid(113 mg, 100%).

Step 2:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenylcarbamoyl)-4-methylthiazol-5-yl)piperazine-1-carboxylate(113 mg, 0.19 mmol) in DCM (2 ml) at 0° C. was added TFA (2 ml)dropwise. And the mixture was stirred at room temperature for 2 h. Thereaction mixture was concentrated in vacuum. The residue was purified byPrep-HPLC to afford a yellow solid compound 042 (52 mg, 71.5%). LCMS:m/z=384.2 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 7.61 (s, 2H), 7.32 (d, J=7.8Hz, 1H), 6.82 (d, J=8.3 Hz, 1H), 6.60 (s, 1H), 6.51 (s, 1H), 5.15 (s,2H), 2.85 (s, 7H), 2.35 (s, 4H).

Example 43—Synthesis of Compound 043

Step 1:

To a solution of 5 lithium5-(4-(tert-butoxycarbonyl)piperazin-1-yl)-4-methylthiazole-2-carboxylate(100 mg, 0.31 mmol) and HATU (116 mg, 0.31 mmol) in DMF (4 ml) wasstirred at room temperature for 15 min. Tert-butyl2-amino-4-(furan-3-yl)phenylcarbamate (47 mg, 0.17 mmol) was added. Themixture was stirred at room temperature for overnight. The residue wasadded water (˜5 mL) and the precipitate was afforded as a yellow solid(73 mg, 73%).

Step 2:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl)phenylcarbamoyl)-4-methylthiazol-5-yl)piperazine-1-carboxylate(73 mg, 0.13 mmol) in DCM (2 ml) at 0° C. was added TFA (2 ml) dropwise.The mixture was stirred at room temperature for 2 h. The reactionmixture was concentrated in vacuum. The residue was purified byPrep-HPLC to afford a yellow solid (45 mg, 93.8%). LCMS: m/z=384.3(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.85 (s, 1H), 8.15 (d, J=19.5 Hz, 1H),7.95 (s, 1H), 7.67 (s, 1H), 7.46 (s, 1H), 7.24 (d, J=8.0 Hz, 1H), 6.80(d, J=5.4 Hz, 2H), 5.00 (s, 2H), 3.13 (s, 4H), 3.02 (s, 4H), 2.37 (s,3H).

Example 44—Synthesis of Compound 044

Step 1:

A mixture of methyl 2-bromothiazole-5-carboxylate (2.22 g, 10 mmol),tert-butyl piperazine-1-carboxylate (3.72 g, 20 mmol) and DIPEA (3.87 g,30 mmol) in 1,4-dioxane (20 mL) was stirred at 100° C. for overnight.The mixture was concentrated to get a residue, which was purified bysilica gel to get desired compound (2.94 g, 90%).

Step 2:

A mixture of methyl2-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-5-carboxylate (327 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 2-amino-4-(pyridin-2-yl)phenylcarbamate (285 mg, 1 mmol),EDCI (573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to get desired compound (435 mg, 75%).

Step 3:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-2-yl)phenylcarbamoyl)thiazol-2-yl)piperazine-1-carboxylate(290 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to afford compound 044 (124 mg, 65%). LCMS: m/z=381.2 (M+H)⁺.¹H NMR (400 MHz, DMSO) δ 9.63 (s, 1H), 8.54 (d, J=4.7 Hz, 1H), 8.07 (s,1H), 7.91 (s, 1H), 7.80-7.69 (m, 3H), 7.18 (t, J=6.5 Hz, 1H), 6.83 (d,J=8.5 Hz, 1H), 5.28 (s, 2H), 3.44-3.38 (m, 4H), 2.82-2.75 (m, 4H).

Example 45—Synthesis of Compound 045

Step 1:

A mixture of methyl2-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-5-carboxylate (327 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 2-amino-4-(pyridin-3-yl)phenylcarbamate (285 mg, 1 mmol),EDCI (573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to afford desired compound (435 mg, 75%).

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-3-yl)phenylcarbamoyl)thiazol-2-yl)piperazine-1-carboxylate(290 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to afford compound 045 (124 mg, 65%). LCMS: m/z=381.2 (M+H)⁺.¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.79 (s, 1H), 8.44 (d, J=4.4 Hz,1H), 8.05 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.50 (d, J=1.8 Hz, 1H), 7.39(td, J=8.3, 3.4 Hz, 2H), 6.88 (d, J=8.3 Hz, 1H), 5.19 (s, 2H), 3.45-3.37(m, 4H), 2.79 (s, 4H).

Example 46—Synthesis of Compound 046

Step 1:

A mixture of methyl2-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-5-carboxylate (327 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 2-amino-4-(furan-3-yl)phenylcarbamate (274 mg, 1 mmol), EDCI(573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to afford desired compound (398 mg, 70%).

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl)phenylcarbamoyl)thiazol-2-yl)piperazine-1-carboxylate(285 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to afford compound 046 (120 mg, 65%) LCMS: m/z=370.4 (M+H)⁺.¹H NMR (400 MHz, DMSO) δ 9.57 (s, 2H), 8.05 (s, 2H), 7.97 (s, 2H), 7.66(s, 2H), 7.32 (s, 2H), 7.22 (d, J=7.8 Hz, 2H), 6.81 (s, 2H), 6.77 (d,J=8.2 Hz, 2H), 3.44 (s, 4H), 2.89 (s, 4H).

Example 47—Synthesis of Compound 047

Step 1:

A mixture of methyl2-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-5-carboxylate (327 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 3-aminobiphenyl-4-ylcarbamate (284 mg, 1 mmol), EDCI (573 mg,3 mmol) and Py (10 mL). The mixture was stirred at room temperature forovernight. The mixture was poured into water (50 mL), filtered to afforddesired compound (405 mg, 70%).

Step 2:

To a solution of tert-butyl4-(5-(4-(tert-butoxycarbonylamino)biphenyl-3-ylcarbamoyl)thiazol-2-yl)piperazine-1-carboxylate(290 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to afford compound 047 (133 mg, 70%). LCMS: m/z=380.1 (M+H)⁺.¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.79 (s, 1H), 8.44 (d, J=4.4 Hz,1H), 8.34 (m, 1H), 8.05 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.50 (d, J=1.8Hz, 1H), 7.39 (td, J=8.3, 3.4 Hz, 2H), 6.88 (d, J=8.3 Hz, 1H), 5.19 (s,2H), 3.45-3.37 (m, 4H), 2.79 (s, 4H).

Example 48—Synthesis of Compound 048

Step 1:

A mixture of methyl 2-bromothiazole-5-carboxylate (2.22 g, 10 mmol),tert-butyl piperazine-1-carboxylate (2.00 g, 20 mmol) and DIPEA (3.87 g,30 mmol) in 1,4-dioxane (20 mL) was stirred at 100° C. for overnight.The mixture was concentrated to get a residue, which was purified bysilica gel to afford desired compound (2.17 g, 90%)

Step 2:

A mixture of methyl 2-(4-methylpiperazin-1-yl)thiazole-5-carboxylate(241 mg, 1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirredat 60° C. for 3 h. The mixture was concentrated to get a residue, whichwas added tert-butyl 3-aminobiphenyl-4-ylcarbamate (284 mg, 1 mmol),EDCI (573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to get desired compound (371 mg, 75%)

Step 3:

To a solution of tert-butyl3-(2-(4-methylpiperazin-1-yl)thiazole-5-carboxamido)biphenyl-4-ylcarbamate(247 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to afford compound 048 (138 mg, 70%). LCMS: m/z=394.2 (M+H)⁺.¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.79 (s, 1H), 8.44 (d, J=4.4 Hz,1H), 8.34 (m, 1H), 8.05 (s, 1H), 7.94 (d, J=8.0 Hz, 1H), 7.50 (d, J=1.8Hz, 1H), 7.39 (td, J=8.3, 3.4 Hz, 2H), 6.88 (d, J=8.3 Hz, 1H), 5.19 (s,2H), 3.45-3.37 (m, 4H), 2.79 (s, 4H), 2.32 (s, 3H).

Example 49—Synthesis of Compound 049

Step 1:

At 0° C., to a mixture of benzyl 2-aminoacetate hydrochloride (5.00 g,24.80 mmol), and pyridine (6.0 mL, 74.39 mmol) in DCM (100 mL) was addedmethyl 2-chloro-2-oxoacetate (2.51 mL, 27.28 mmol). It was stirred at 0°C. to room temperature for 18 h. Then it was washed with water andconcentrated in vacuo. The residue was washed with EA-PE to give crudecompound (4.73 g).

Step 2:

Under H₂, a mixture of methyl2-(2-(benzyloxy)-2-oxoethylamino)-2-oxoacetate (8.55 g, 34.03 mmol) andPd/C (10%, 700 mg) in MeOH (150 mL) was stirred at room temperature for18 h. After filtering, the filtrate was concentrated in vacuo to givecompound as a white solid (5.54 g, yield: 100%).

Step 3:

A mixture of tert-2-(2-methoxy-2-oxoacetamido)acetic acid (4.38 g, 27.19mmol), EDCI (5.73 g, 29.91 mmol) and HOBT (4.04 g, 29.91 mmol) in DMF(100 mL) was stirred at room temperature for 15 mins. Then DIEA (18 mL,108.76 mmol) and tert-butyl piperazine-1-carboxylate (5.57 g, 29.91mmol) was added. It was stirred at room temperature for 18 h. Theresulting precipitate was filtered to give the compound (1.80 g, yield:20%) as a white solid.

Step 4:

Under N2, a mixture of tert-butyl4-(2-(2-methoxy-2-oxoacetamido)acetyl)piperazine-1-carboxylate (100 mg,0.30 mmol), phosphorus pentasulfide (100 mg, 0.23 mmol) in pyridine (10mL) was stirred at 100° C. for 4 h. It was cooled to room temperatureand concentrated in vacuo. The residue was purified by prep-TLC to givecompound as a light yellow solid (47 mg, yield: 47%).

Step 5:

At 0° C., a mixture of methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-2-carboxylate 128 mg,0.39 mmol) and LiOH (33 mg, 0.78 mmol) in THF (6 ml) MeOH (1 mL) andwater (2 mL) was stirred at 0° C. to room temperature for 2 h. It wasconcentrated in vacuo to give crude compound as a Li salt.

Step 6:

A mixture of above Li salt (0.39 mmol) and HATU (164 mg, 0.43 mol) inDMF (4 mL) was stirred at room temperature for 15 mins. Tert-butyl3-aminobiphenyl-4-ylcarbamate (122 mg, 0.43 mmol) was added. It wasstirred at room temperature for 18 h. Water was added. The precipitatewas collected by filtration to give crude compound as a brown solid.

Step 7:

At 0° C., to a solution of above tert-butyl4-(2-(4-(tert-butoxycarbonylamino)biphenyl-3-ylcarbamoyl)thiazol-5-yl)piperazine-1-carboxylate(0.39 mmol) in DCM (2 mL) was added TFA (2 mL). It was stirred at 0° C.for 2 h. Then it was concentrated in vacuo. The residue was purified byPrep-HPLC to compound 049 as a yellow solid (51.3 mg, yield: 36%, threesteps). LCMS: m/z=380.2 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.73 (s, 1H),7.64 (s, 1H), 7.54 (d, J=7.7 Hz, 2H), 7.39 (t, J=7.6 Hz, 2H), 7.28 (dd,J=15.2, 6.8 Hz, 3H), 6.87 (d, J=8.3 Hz, 1H), 5.09 (s, 2H), 3.15 (s, 4H),2.84 (s, 4H).

Example 50—Synthesis of Compound 050

Step 1:

A mixture of above Li salt (0.39 mmol) and HATU (164 mg, 0.43 mol) inDMF (4 mL) was stirred at room temperature for 15 mins. Tert-butyl2-amino-4-(furan-2-yl)phenylcarbamate (118 mg, 0.43 mmol) was added. Itwas stirred at room temperature for 18 h. Water was added. Theprecipitate was collected by filtration to give crude compound as abrown solid.

Step 2:

At 0° C., to a solution of above tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenylcarbamoyl)thiazol-5-yl)piperazine-1-carboxylate(0.39 mmol) in DCM (2 mL) was added TFA (2 mL). It was stirred at 0° C.for 2 h. Then it was concentrated in vacuo. The residue was purified byPrep-HPLC to compound 050 as a yellow solid (51.3 mg, yield: 36%, threesteps). LCMS: m/z=370.0 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.67 (s, 1H),7.65 (d, J=2.0 Hz, 1H), 7.61 (s, 1H), 7.30 (dd, J=8.4, 2.0 Hz, 1H), 7.25(s, 1H), 6.81 (d, J=8.0 Hz, 1H), 6.61 (d, J=2.8 Hz, 1H), 6.51 (m, 1H),5.14 (s, 2H), 3.16 (m, 4H), 2.84 (m, 4H).

Example 51—Synthesis of Compound 051

Step 1:

A mixture of above Li salt (0.42 mmol) and HATU (175 mg, 0.46 mol) inDMF (4 mL) was stirred at room temperature for 15 mins. Tert-butyl2-amino-4-(furan-3-yl)phenylcarbamate (126 mg, 0.46 mmol) was added. Itwas stirred at room temperature for 4 h. Water was added. Theprecipitate was collected by filtration. It was purified by prep-TLC togive compound (158 mg, yield: 64%) as a light yellow solid.

Step 2:

At 0° C., to a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl)phenylcarbamoyl)thiazol-5-yl)piperazine-1-carboxylate (153 mg, 0.27mmol) in DCM (2 mL) was added TFA (2 mL). It was stirred at 0° C. for 2h. Then it was concentrated in vacuo. The residue was neutralized withaqueous saturated NaHCO₃ to pH8. The resulting solid was collected anddried in vacuo to compound 051 as a light yellow solid (31.7 mg, yield:32%). LCMS: m/z=369.9 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.67 (s, 1H),7.96 (s, 1H), 7.67 (m, 1H), 7.51 (d, J=1.6 Hz, 1H), 7.24 (s, 1H), 7.22(dd, J=8.4, 2.0 Hz, 1H), 6.80 (m, 2H), 4.97 (s, 2H), 3.16 (m, 4H), 2.84(m, 4H).

Example 52—Synthesis of Compound 052

Step 1:

A mixture of5-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-2-carboxylic acid (120mg, 0.38 mmol), tert-butyl 2-amino-4-(pyridin-4-yl)phenylcarbamate (120mg, 0.42 mmol) and EDCI (147 mg, 0.77 mmol) in pyridine (6 mL) wasstirred at room temperature for 18 h. Then it was concentrated in vacuoand the residue was washed with water and purified by prep-TLC to givecompound (140 mg, 63%) as a yellow solid.

Step 2:

At 0° C., to a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(pyridine-4-yl)phenylcarbamoyl)thiazol-5-yl)piperazine-1-carboxylate(140 mg, 0.24 mmol) in DCM (2 mL) was added TFA (2 mL). It was stirredat 0° C. for 2 h. Then it was concentrated in vacuo. The residue wasneutralized with aqueous saturated NaHCO₃ to pH8. The resulting solidwas collected and dried in vacuo to compound 052 as a light yellow solid(77.1 mg, yield: 84%). LCMS: m/z=381.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ9.76 (s, 1H), 8.51 (dd, J=4.6, 1.5 Hz, 2H), 7.77 (d, J=2.1 Hz, 1H), 7.57(dd, J=4.7, 1.6 Hz, 2H), 7.48 (dd, J=8.4, 2.2 Hz, 1H), 7.25 (s, 1H),6.89 (d, J=8.4 Hz, 1H), 5.36 (s, 2H), 3.18-3.12 (m, 4H), 2.89-2.79 (m,4H).

Example 53—Synthesis of Compound 053

Step 1:

A mixture of5-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-2-carboxylic acid (60mg, 0.19 mmol), tert-butyl 3-amino-4′-fluorobiphenyl-4-ylcarbamate (64mg, 0.21 mmol) and EDCI (73 mg, 0.38 mmol) in pyridine (6 mL) wasstirred at room temperature for 18 h. Then it was concentrated in vacuoand the residue was washed with water and purified by prep-TLC to givecompound (140 mg, 63%) as a white solid.

Step 2:

At 0° C., to a solution of tert-butyl4-(2-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)thiazol-5-yl)piperazine-1-carboxylate(140 mg, 0.23 mmol) in DCM (2 mL) was added TFA (2 mL). It was stirredat 0° C. for 4 h. Then it was concentrated in vacuo. The residue wasneutralized with aqueous saturated NaHCO₃ to pH8. The resulting solidwas collected and dried in vacuo to compound 53 as a light yellow solid(17.0 mg, yield: 18%). LCMS: m/z=398.2 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ9.72 (s, 1H), 7.69-7.50 (m, 3H), 7.34-7.16 (m, 4H), 6.86 (d, J=8.3 Hz,1H), 5.09 (s, 2H), 3.21-3.09 (m, 4H), 2.90-2.77 (m, 4H).

Example 54—Synthesis of Compound 054

Step 1:

A mixture of5-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-2-carboxylic acid (153mg, 0.49 mmol) and HATU (204 mg, 0.54 mmol) in DMF (2 mL) was stirred atroom temperature for 15 mins. Then DIEA (188 mg, 1.46 mmol) andtert-butyl 2-amino-4-(furan-2-yl)phenylcarbamate (147 mg, 0.54 mmol)were added. It was stirred at room temperature for 1 h. Then water wasadded. The resulting solid was collected by filtration and dried invacuo to give compound.

Step 2:

To a mixture of above tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenylcarbamoyl)thiazol-5-yl)piperazine-1-carboxylate in DCM (2 mL) was addedTFA (2 mL) at 0° C. It was stirred at 0° C. to room temperature for 2 h.It was concentrated in vacuo and the residue was washed with aqueoussaturated NaHCO₃ solution to give product (160 mg) as a white solid.

Step 3:

It was stirred at 0° C. to give a complete reaction. It was concentratedin vacuo and the residue was purified by Prep-HPLC to give compound 054as a yellow solid. LCMS: m/z=384.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.67(s, 1H), 7.63 (d, J=17.9 Hz, 2H), 7.30 (d, J=6.8 Hz, 1H), 7.26 (s, 1H),6.82 (d, J=8.7 Hz, 1H), 6.60 (d, J=3.3 Hz, 1H), 6.51 (s, 1H), 5.13 (s,2H), 3.28-3.23 (m, 4H), 2.46 (d, J=5.1 Hz, 4H), 2.23 (s, 3H).

Example 55—Synthesis of Compound 055

Step 1:

A mixture of 4-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiazole-2-carboxylic acid (48 mg, 0.15 mmol), tert-butyl2-amino-4-(furan-2-yl) phenylcarbamate (42 mg, 0.15 mmol) and EDCI (88mg, 0.46 mmol) in Py (3 ml) was stirred at room temperature forovernight. It was purified by column chromatography (DCM:EA=5:1). Lightyellow solid was afforded (50 mg, 57% yield).

Step 2:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl) phenylcarbamoyl)thiazol-4-yl) piperazine-1-carboxylate (50 mg, 0.09 mmol) in DCM (3 ml)was added HCl/1,4-dioxane (1 ml). The mixture was stirred at roomtemperature for 2 h. It was concentrated and washed by Et₂O. Yellowsolid was afforded as compound 055. (16 mg, HCl salt, lotSP-0017550-050). LCMS: m/z=370.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 10.23(s, 1H), 9.24 (s, 2H), 7.70 (t, J=13.0 Hz, 2H), 7.50 (d, J=8.0 Hz, 1H),7.13 (s, 1H), 6.88 (d, J=14.2 Hz, 1H), 6.77 (d, J=28.9 Hz, 1H), 6.57 (d,J=1.8 Hz, 1H), 3.63 (s, 4H), 3.24 (s, 4H).

Example 56—Synthesis of Compound 056

Step 1:

A mixture of 4-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiazole-2-carboxylic acid (59 mg, 0.19 mmol), tert-butyl2-amino-4-(furan-3-yl) phenylcarbamate (52 mg, 0.19 mmol) and EDCI (109mg, 0.57 mmol) in Py (3 ml) was stirred at room temperature forovernight. It was purified by column chromatography (DCM:EA=5:1). Lightyellow solid was afforded (25 mg, 23% yield).

Step 2:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl) phenylcarbamoyl)thiazol-4-yl) piperazine-1-carboxylate (25 mg, 0.04 mmol) in DCM (2 ml)was added TFA (1 ml). The mixture was stirred at room temperature for 2h. It was concentrated and washed by Et₂O. Yellow solid was afforded asCompound 056. (20 mg, TFA salt, lot SP-0018270-093). LCMS: m/z=370.1(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.64 (s, 1H), 9.37 (s, 2H), 8.28 (s,1H), 7.87-7.76 (m, 2H), 7.60 (dd, J=8.4, 1.7 Hz, 1H), 7.48 (s, 1H), 6.93(s, 1H), 6.87-6.78 (m, 1H), 4.39 (s, 2H), 3.93 (d, J=57.2 Hz, 4H), 3.58(s, 1H), 3.27 (s, 3H).

Example 57—Synthesis of Compound 057

Step 1:

A mixture of 4-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiazole-2-carboxylic acid (45 mg, 0.14 mmol), tert-butyl2-amino-4-(pyridin-4-yl) phenylcarbamate (83 mg, 0.14 mmol) and EDCI (83mg, 0.43 mmol) in Py (3 ml) was stirred at room temperature forovernight. It was purified by column chromatography (DCM:EA=1:1). Greensolid was afforded (40 mg, 48% yield).

Step 2:

To a solution of tert-butyl4-(2-(2-(tert-butoxycarbonylamino)-5-(pyridin-4-yl) phenylcarbamoyl)thiazol-4-yl) piperazine-1-carboxylate (40 mg, 0.07 mmol) in DCM (2 ml)was added HCl/1,4-dioxane (1 ml). The mixture was stirred at roomtemperature for 2 h. It was concentrated and washed by Et₂O. Yellowsolid was afforded as compound 057. (20 mg, HCl salt, lotSP-0017550-067). LCMS: m/z=381.0 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 10.02(s, 1H), 9.48 (s, 2H), 8.71 (d, J=6.4 Hz, 2H), 8.19 (d, J=6.5 Hz, 2H),7.97 (d, J=1.9 Hz, 1H), 7.87-7.79 (m, 1H), 6.94 (d, J=8.6 Hz, 1H), 6.88(s, 1H), 3.63 (s, 4H), 3.23 (s, 4H).

Example 58—Synthesis of Compound 058

Step 1:

A mixture of 4-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiazole-2-carboxylic acid (45 mg, 0.14 mmol), tert-butyl3-amino-4′-fluorobiphenyl-4-ylcarbamate (86 mg, 0.14 mmol) and EDCI (83mg, 0.43 mmol) in Py (3 ml) was stirred at room temperature forovernight. It was extracted by EA and purified by column chromatography(DCM:EA=5:1). Yellow solid was afforded (30 mg, 35% yield).

Step 2:

To a solution of tert-butyl4-(2-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)thiazol-4-yl) piperazine-1-carboxylate (30 mg, 0.05 mmol) in DCM (2 ml)was added HCl/1,4-dioxane (1 ml). The mixture was stirred at roomtemperature for 2 h. It was concentrated and washed by Et₂O. Yellowsolid was afforded as compound 058. (22 mg, HCl salt, lotSP-0017550-058). LCMS: m/z=398.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 10.37(s, 1H), 9.30 (s, 2H), 7.71 (d, J=31.3 Hz, 3H), 7.51 (s, 1H), 7.29 (t,J=8.8 Hz, 3H), 6.91 (s, 1H), 3.60 (d, J=30.0 Hz, 4H), 3.24 (s, 4H).

Example 59—Synthesis of Compound 059

Step 1:

Tert-butyl piperazine-1-carboxylate (1.8 g, 10 mmol), ethyl2-bromothiazole-4-carboxylate (2.36 g, 10 mmol) and DIPEA (2.02 g, 20mmol) were added into 1,4-dioxane (15 ml). The mixture was stirred forovernight at 100° C. When the reaction finished, it was extracted by EAand washed by brine. It was purified by flash column chromatography(PE/EA=5:1-3:1) to afford the yellow solid as product (3.0 g, 88%).

Step 2:

Ethyl 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-4-carboxylate(2.0 g) was added into a solution of EtOH (10 ml), THF (10 ml) and NaOH(2 N, 5 ml). The mixture was stirred at 60° C. for 2 h. When thereaction finished, the pH was adjusted to 1 with 1N HCl, the precipitatewas collected, dried to afford the pale solid (1.7 g, 94 yield).

Step 3:

2-(4-(tert-butoxycarbonyl)piperazin-1-yl)thiazole-4-carboxylic acid (156mg, 0.5 mmol), tert-butyl 2-amino-4-(thiophen-2-yl) phenylcarbamate (130mg, 0.45 mmol) and EDCI (191 mg, 1 mmol) were added into Py (5 ml). Themixture was stirred at room temperature for overnight. When the reactionfinished, it was concentrated and water was added. The yellow solid wascollected as product (210 mg, 80% yield).

Step 4:

Tert-butyl4-(4-(2-(tert-butoxycarbonylamino)-5-(thiophen-2-yl)phenylcarbamoyl)thiazol-2-yl)piperazine-1-carboxylate(210 mg, 0.36 mmol) was dissolved into DCM (5 ml). Then TFA (1 ml) wasadded. The mixture was stirred at room temperature for 2 h. When thereaction finished, it was concentrated and ether was added to affordcompound 059 as a yellow solid product (150 mg, TFA salt, LotsSP-008851-024). LCMS: m/z=386.5 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.72(s, 1H), 8.47 (s, 1H), 7.70 (J=8.0 Hz, 1H), 7.54 (s, 1H), 7.40 (d, J=8.0Hz, 1H), 7.13 (m, 3H), 6.99 (dd, J=8.8 Hz, 1H), 6.67 (d, J=8.8 Hz, 1H),3.43 (m, 4H), 2.81 (m, 4H).

Example 60—Synthesis of Compound 060

Step 1:

Tert-butyl piperazine-1-carboxylate (744 mg, 4 mmol), ethyl2-chlorooxazole-5-carboxylate (700 mg, 4 mmol) and DIPEA (1032 mg, 8mmol) were added into 1,4-dioxane (15 ml). The mixture was stirred for 2h at 100° C. When the reaction finished, it was cooled and water wasadded. The precipitate was collected to afford the yellow solid asproduct (1.45 g, crude).

Step 2:

Ethyl 2-(4-(tert-butoxycarbonyl)piperazin-1-yl)oxazole-5-carboxylate(1.45 g, crude) was added into a solution of EtOH (5 ml), THF (5 ml) andNaOH (2 N, 5 ml). The mixture was stirred at 60° C. for 2 h. When thereaction finished, the pH was adjusted to 1. The solid was collected toafford the pale solid. (1.3 g, 91% yield, 2 steps).

Step 3:

2-(4-(tert-butoxycarbonyl)piperazin-1-yl)oxazole-5-carboxylic acid (300mg, 1 mmol), tert-butyl 2-amino-4-(thiophen-2-yl) phenylcarbamate (261mg, 0.9 mmol) and EDCI (382 mg, 2 mmol) were added into Py (8 ml). Themixture was stirred at room temperature for overnight. When the reactionfinished, it was concentrated and washed with water. The pale yellowsolid was collected as product (600 mg, 100% yield).

Step 4:

Tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(thiophen-2-yl)phenylcarbamoyl)oxazol-2-yl)piperazine-1-carboxylate(362 mg, crude) was dissolved into DCM (5 ml). Then TFA (2 ml) wasadded. The mixture was stirred at room temperature for 2 h. When thereaction finished, it was adjusted to pH=10 and extracted by EA. It wasconcentrated and washed by 5 ml EA to afford pure compound 060 as awhite solid product (78 mg, 38% yield, Lots SP-0017054-174). LCMS:m/z=369.4 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.42 (s, 1H), 7.62 (s, 1H),7.44-7.33 (m, 2H), 7.27 (dd, J=19.5, 5.6 Hz, 2H), 7.12-6.98 (m, 1H),6.80 (d, J=8.4 Hz, 1H), 5.14 (s, 2H), 5.14 (s, 2H), 3.48 (s, 4H), 3.48(s, 4H), 2.78 (s, 4H).

Example 61—Synthesis of Compound 061

Step 1:

2-(4-(tert-butoxycarbonyl)piperazin-1-yl)oxazole-5-carboxylic acid (205mg, 0.69 mmol), tert-butyl 2-amino-4-(furan-2-yl)phenylcarbamate (170mg, 0.621 mmol) and EDCI (267 mg, 1.4 mmol) were added into Py (8 ml).The mixture was stirred at room temperature for overnight. When thereaction finished, it was concentrated and washed with water. The paleyellow solid was collected as product (360 mg, 100% yield).

Step 2:

Tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenylcarbamoyl)oxazol-2-yl)piperazine-1-carboxylate(360 mg, crude) was dissolved into DCM (5 ml). Then TFA (2 ml) wasadded. The mixture was stirred at room temperature for 2 h. When thereaction finished, it was adjusted to pH=10 and extracted with EA. Itwas concentrated and washed with 5 ml EA to afford pure compound 061 asa white solid product (109 mg, 53% yield, Lots SP-0017054-180). LCMS:m/z=354.4 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.45 (s, 1H), 7.74-7.54 (m,2H), 7.45 (d, J=1.9 Hz, 1H), 7.32 (dd, J=8.3, 2.0 Hz, 1H), 6.80 (d,J=8.4 Hz, 1H), 6.61 (d, J=3.3 Hz, 1H), 6.49 (dt, J=22.4, 11.2 Hz, 1H),5.11 (d, J=30.3 Hz, 2H), 3.55-3.43 (m, 4H), 2.87-2.71 (m, 4H).

Example 62—Synthesis of Compound 062

Step 1:

2-(4-(tert-butoxycarbonyl)piperazin-1-yl)oxazole-5-carboxylic acid (205mg, 0.69 mmol), tert-butyl 3-aminobiphenyl-4-ylcarbamate (176 mg, 0.62mmol) and EDCI (267 mg, 1.4 mmol) were added into Py (8 ml). The mixturewas stirred at room temperature for overnight. When the reactionfinished, it was concentrated and washed with water. The pale yellowsolid was collected as product (893 mg, crude).

Step 2:

Tert-butyl4-(5-(4-(tert-butoxycarbonylamino)biphenyl-3-ylcarbamoyl)oxazol-2-yl)piperazine-1-carboxylate(362 mg, crude) was dissolved into DCM (5 ml). Then TFA (2 ml) wasadded. The mixture was stirred at room temperature for 2 h. When thereaction finished, it was adjusted to pH=10 and extracted with EA. Itwas concentrated and washed with 5 ml EA to afford pure compound 062 asa white solid product (170 mg, 68% yield, Lots SP-0017054-177). LCMS:m/z=364.4 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.45 (s, 1H), 7.70-7.16 (m,9H), 6.86 (d, J=8.1 Hz, 1H), 5.09 (s, 2H), 3.46 (d, J=28.8 Hz, 4H), 2.79(s, 4H).

Example 63—Synthesis of Compound 063

Step 1:

2-(4-(tert-butoxycarbonyl)piperazin-1-yl)oxazole-5-carboxylic acid (155mg, 0.52 mmol), tert-butyl 3-amino-4′-fluorobiphenyl-4-ylcarbamate (141mg, 0.47 mmol) and HATU (300 mg, 0.78 mmol), DIPEA (134 mg, 1.04 mmol)were added into THF (8 ml). The mixture was stirred at room temperaturefor overnight. When the reaction finished, it was purified on silica gelcolumn to afford the product (202 mg, 96%).

Step 2:

Tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)oxazol-2-yl)piperazine-1-carboxylate(202 mg, 0.35 mmol) was dissolved into DCM (5 ml). Then TFA (2 ml) wasadded. The mixture was stirred at room temperature for 2 h. When thereaction finished, it was adjusted to pH=10 and extracted with EA toafford pure compound 063 as a white solid product (123 mg, 92% yield,Lots SP-0017054-188). LCMS: m/z=382.4 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ9.45 (s, 1H), 7.64 (s, 1H), 7.57 (dd, J=8.6, 5.5 Hz, 2H), 7.42 (d, J=1.7Hz, 1H), 7.29 (dd, J=8.3, 2.0 Hz, 1H), 7.21 (t, J=8.8 Hz, 2H), 6.85 (d,J=8.3 Hz, 1H), 5.09 (s, 2H), 3.53-3.43 (m, 4H), 2.86-2.73 (m, 4H).

Example 64—Synthesis of Compound 064

Step 1:

A mixture of 5-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiophene-2-carboxylic acid (94 mg, 0.3 mmol), tert-butyl3-aminobiphenyl-4-ylcarbamate (85 mg, 0.3 mmol) and EDCI (173 mg, 0.9mmol) in Py (3 ml) was stirred at room temperature for overnight. It wasextracted with EA and purified by column chromatography (PE:EA=2:1).Green solid was afforded (140 mg, 61%).

Step 2:

To a solution of tert-butyl4-(5-(4-(tert-butoxycarbonylamino)biphenyl-3-ylcarbamoyl)thiophen-2-yl)piperazine-1-carboxylate(120 mg, 0.21 mmol) in DCM (2 ml) was added TFA (2 ml). The mixture wasstirred at room temperature for 2 h. It was concentrated and washed withEt₂O. Yellow solid was afforded as compound 064. (130 mg, TFA salt, lotSP-0017550-024). LCMS: m/z=379.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.64(s, 1H), 8.91 (s, 2H), 7.80 (d, J=4.2 Hz, 1H), 7.56 (d, J=7.3 Hz, 2H),7.49 (d, J=1.9 Hz, 1H), 7.44-7.33 (m, 3H), 7.27 (t, J=7.4 Hz, 1H), 6.94(d, J=8.3 Hz, 1H), 6.36 (d, J=4.2 Hz, 1H), 3.42 (d, J=5.7 Hz, 4H), 3.28(s, 4H).

Example 65—Synthesis of Compound 065

Step 1:

A mixture of 5-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiophene-2-carboxylic acid (66 mg, 0.2 mmol), tert-butyl2-amino-4-(furan-2-yl) phenylcarbamate (58 mg, 0.2 mmol) and EDCI (121mg, 0.6 mmol) in Py (2 ml) was stirred at room temperature forovernight. It was extracted with EA and purified by columnchromatography (PE:EA=2:1). White solid was afforded (50 mg, 42%).

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl) phenylcarbamoyl)thiophen-2-yl) piperazine-1-carboxylate (50 mg, 0.09 mmol) in DCM (2 ml)was added TFA (1 ml). The mixture was stirred at room temperature for 2h. It was concentrated and washed with Et₂O. White solid was afforded ascompound 065. (60 mg, TFA salt, lot SP-0017550-034). LCMS: m/z=369.1(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.51 (s, 1H), 8.94 (s, 2H), 7.77 (d,J=4.2 Hz, 1H), 7.61 (s, 1H), 7.47 (d, J=1.9 Hz, 1H), 7.32 (dd, J=8.3,1.9 Hz, 1H), 6.81 (d, J=8.4 Hz, 1H), 6.61 (d, J=3.2 Hz, 1H), 6.51 (dd,J=3.2, 1.8 Hz, 1H), 6.35 (d, J=4.2 Hz, 1H), 3.42 (d, J=5.4 Hz, 4H), 3.27(s, 4H).

Example 66—Synthesis of Compound 066

Step 1:

A mixture of 5-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiophene-2-carboxylic acid (78 mg, 0.25 mmol), tert-butyl2-amino-4-(furan-3-yl) phenylcarbamate (69 mg, 0.25 mmol) and EDCI (144mg, 0.75 mmol) in Py (2 ml) was stirred at room temperature forovernight. It was extracted with EA and purified by columnchromatography (PE:EA=2:1). White solid was afforded (110 mg, 77%).

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl) phenylcarbamoyl)thiophen-2-yl) piperazine-1-carboxylate (110 mg, 0.19 mmol) in DCM (2ml) was added TFA (1 ml). The mixture was stirred at room temperaturefor 2 h. It was concentrated and washed with Et₂O. White solid wasafforded as compound 066. (110 mg, TFA salt, lot SP-0017550-031). LCMS:m/z=369.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.63 (s, 1H), 8.89 (s, 2H),8.00 (s, 1H), 7.78 (d, J=4.1 Hz, 1H), 7.68 (t, J=1.6 Hz, 1H), 7.37 (d,J=1.8 Hz, 1H), 7.28 (d, J=8.3 Hz, 1H), 6.92-6.78 (m, 2H), 6.35 (d, J=4.3Hz, 1H), 3.41 (d, J=5.2 Hz, 4H), 3.28 (s, 4H).

Example 67—Synthesis of Compound 067

Step 1:

A mixture of 5-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiophene-2-carboxylic acid (66 mg, 0.21 mmol), tert-butyl2-amino-4-(pyridin-4-yl)phenylcarbamate (60 mg, 0.21 mmol) and EDCI (121mg, 0.63 mmol) in Py (3 ml) was stirred at room temperature forovernight. It was extracted with EA and concentrated for next step (100mg, crude).

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-4-yl) phenylcarbamoyl)thiophen-2-yl) piperazine-1-carboxylate (100 mg, 0.17 mmol) in DCM (2ml) was added TFA (1 ml). The mixture was stirred at room temperaturefor 2 h. It was concentrated and purified by prep-HPLC (base method).White solid was afforded as compound 067. (30 mg, 65% yield, lotSP-0017550-039). LCMS: m/z=380.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.40(s, 1H), 8.50 (t, J=8.5 Hz, 2H), 7.75 (d, J=4.2 Hz, 1H), 7.63 (d, J=2.0Hz, 1H), 7.58 (d, J=6.1 Hz, 2H), 7.48 (dd, J=8.3, 2.0 Hz, 1H), 6.88 (d,J=8.4 Hz, 1H), 6.19 (t, J=6.6 Hz, 1H), 5.32 (s, 2H), 3.15-3.07 (m, 4H),2.87-2.77 (m, 4H).

Example 68—Synthesis of Compound 068

Step 1:

A mixture of 5-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiophene-2-carboxylic acid (70 mg, 0.22 mmol), tert-butyl3-amino-4′-fluorobiphenyl-4-ylcarbamate (68 mg, 0.22 mmol) and EDCI (129mg, 0.67 mmol) in Py (3 ml) was stirred at room temperature forovernight. It was extracted with EA and purified by columnchromatography (PE:EA=2:1). White solid was afforded (120 mg, 87%yield).

Step 2:

To a solution of tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)thiophen-2-yl) piperazine-1-carboxylate (120 mg, 0.2 mmol) in DCM (4 ml)was added TFA (2 ml). The mixture was stirred at room temperature for 2h. It was concentrated and washed with Et₂O. White solid was afforded ascompound 068. (84 mg, TFA salt, lot SP-0017550-044). LCMS: m/z=397.1(M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.58 (s, 1H), 9.06 (s, 2H), 7.79 (d,J=4.0 Hz, 1H), 7.58 (dd, J=8.4, 5.5 Hz, 2H), 7.45 (s, 1H), 7.30 (d,J=6.6 Hz, 1H), 7.22 (t, J=8.8 Hz, 2H), 6.89 (d, J=8.3 Hz, 1H), 6.35 (d,J=4.1 Hz, 1H), 3.42 (d, J=4.9 Hz, 4H), 3.27 (s, 4H).

Example 69—Synthesis of Compound 069

Step 1:

A mixture of methyl 4-bromothiophene-2-carboxylate (220 mg, 1 mmol),tert-butyl piperazine-1-carboxylate (372 mg, 2 mmol), Pd₂(dba)₃ (92 mg,0.1 mmol), RuPhos (47 mg, 0.1 mmol) and Cs₂CO₃ (652 mg, 2 mmol) in PhMe(10 ml) was stirred at 95° C. for overnight under N₂ atmosphere. It wasextracted with EA and purified by column chromatography (PE:EA=5:1).Yellow solid was afforded (360 mg, 92%).

Step 2:

To a solution of tert-butyl 4-(5-(methoxycarbonyl) thiophen-3-yl)piperazine-1-carboxylate (60 mg, 0.18 mmol) and LiOH H₂O (16 mg, 0.37mmol) were added into MeOH (5 ml). The mixture was stirred at 60° C. forovernight. It was concentrated for next step (60 mg, crude).

Step 3:

A mixture of 4-(4-(tert-butoxycarbonyl) piperazin-1-yl)thiophene-2-carboxylic acid (60 mg, 0.19 mmol), tert-butyl3-aminobiphenyl-4-ylcarbamate (55 mg, 0.19 mmol), EDCI (110 mg, 0.58mmol) in Py (3 ml). The mixture was stirred at room temperature forovernight. It was extracted with EA and purified by columnchromatography (PE:EA=2:1). Yellow solid was afforded (90 mg, 81%).

Step 4:

To a solution of tert-butyl 4-(5-(4-(tert-butoxycarbonylamino)biphenyl-3-ylcarbamoyl) thiophen-3-yl) piperazine-1-carboxylate (90 mg,0.16 mmol) in DCM (2 ml) was added TFA (1 ml) dropwise. The mixture wasstirred at room temperature for 2 h. It was concentrated and washed withEt₂O. Grey solid was afforded as compound 069 (67 mg, TFA salt, lotSP-0017550-084). LCMS: m/z=379.1 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.78(s, 1H), 8.84 (s, 2H), 7.91 (s, 1H), 7.56 (d, J=7.3 Hz, 2H), 7.50 (d,J=2.0 Hz, 1H), 7.45-7.33 (m, 3H), 7.26 (t, J=7.3 Hz, 1H), 6.93 (d, J=8.3Hz, 1H), 6.86 (d, J=1.5 Hz, 1H), 3.29 (s, 8H).

Example 70—Synthesis of Compound 070

Step 1:

A mixture of methyl 6-chloronicotinate (500 mg, 2.92 mmol),1-methylpiperazine (900 mg, 9 mmol), DIPEA (774 mg, 6 mmol) in1,4-dioxane (15 mL) was stirred at 100° C. under N₂ atmosphere forovernight. The mixture was cool and EA (50 mL) and water (40 mL) wereadded, stirred for 30 min, the organic layer was separated, dried,concentrated to get a residue, which was washed with PE (30 mL) toafford as yellow solid (650 mg, 88%).

Step 2:

To a solution of methyl 6-(4-methylpiperazin-1-yl)nicotinate (650 mg,2.76 mmol) in THF (10 ml) and EtOH (10 mL) was added 2M NaOH (1.5 mL,3.04 mmol, 1.1 eq). The mixture was stirred at 60° C. for 3 h. When thereaction finished, it was concentrated to remove solvent, HCl (2 M) wasadded to adjust pH 5-7, concentrated to afford crude compound as a whitesolid (1.0 g, crude, contained some NaCl).

Step 3:

A mixture of 6-(4-methylpiperazin-1-yl)nicotinic acid (150 mg, crude),tert-butyl 3-aminobiphenyl-4-ylcarbamate (173 mg, 0.6 mmol) and EDCI(260 mg, 1.35 mmol) in pyridine (10 mL) was stirred at room temperaturefor overnight. The mixture was poured into water (20 mL), filtered toafford desired compound (300 mg, 89%) as a yellow solid.

Step 4:

A mixture of tert-butyl3-(6-(4-methylpiperazin-1-yl)nicotinamido)biphenyl-4-ylcarbamate (300mg, 0.616 mmol) and TFA (2 mL) in DCM (5 mL) was stirred at roomtemperature for 2 h. The mixture was purified by Prep-HPLC to obtaincompound 070 (70 mg, 27%, Lots SP-0016945-112) as a white solid. LCMS:m/z=388.5 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.76 (s, 1H),8.10 (d, J=8.7 Hz, 1H), 7.59-7.48 (m, 3H), 7.32 (ddd, J=35.3, 14.6, 7.2Hz, 4H), 6.89 (dd, J=21.5, 8.6 Hz, 2H), 5.09 (s, 2H), 3.63 (s, 4H), 2.40(s, 4H).

Example 71—Synthesis of Compound 071

Step 1:

A mixture of 6-(4-methylpiperazin-1-yl)nicotinic acid (150 mg, crude),tert-butyl 2-amino-4-(thiophen-2-yl)phenylcarbamate (174 mg, 0.6 mmol)and EDCI (260 mg, 1.35 mmol) in pyridine (10 mL) was stirred at roomtemperature for overnight. The mixture was poured into water (20 mL),filtered to afford desired compound (280 mg, 84%) as a yellow solid.

Step 2:

A mixture of compound 4 (280 mg, 0.5 mmol) and TFA (3 mL) in DCM (3 mL)was stirred at room temperature for 1 h. The mixture was purified byPrep-HPLC to obtain compound 071 (170 mg, 65%, lot SP-0016945-116) as ayellow solid. LCMS: m/z=394.5 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.72 (d,J=12.1 Hz, 1H), 8.81 (t, J=3.6 Hz, 1H), 8.20 (dd, J=9.0, 2.2 Hz, 1H),7.49 (d, J=1.8 Hz, 2H), 7.35 (ddd, J=24.5, 18.3, 4.2 Hz, 4H), 7.07 (dd,J=8.8, 4.9 Hz, 2H), 6.91 (d, J=8.4 Hz, 1H), 4.58 (d, J=13.3 Hz, 2H),3.54 (d, J=11.4 Hz, 2H), 3.20 (dd, J=40.8, 28.4 Hz, 5H), 2.86 (s, 3H).

Example 72—Synthesis of Compound 072

Step 1:

To a solution of 6-(4-(tert-butoxycarbonyl) piperazin-1-yl) nicotinicacid (123.7 mg, 0.40 mmol) and tert-butyl 2-amino-4-(pyridin-3-yl)phenylcarbamate (1, 3, 2-dioxaborolane) (114.5 mg, 0.40 mmol) in Py (2.5ml) was added EDCI (230 mg, 1.20 mmol). The mixture was stirred at roomtemperature for overnight. The residue was purified by preparative TLC(silica gel, GF254 10-40 u, 25×25 cm) with PE/EA (2:1) to afford ayellow solid (165 mg, 72%).

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-3-yl) phenylcarbamoyl)pyridin-2-yl) piperazine-1-carboxylate (165 mg, 0.29 mmol) in DCM (5 ml)at 0° C. was added TFA (2 ml) drop wise. And the resulted was stirred atroom temperature for 2 h. The ice water was added to the resultingmixture and neutralized with saturated NaOH (3 ml) solution. The mixturewas filtered through a Celite pad and the filtrate was concentrated toafford compound 072 as a red solid (40 mg, 39%). LCMS: m/z=375.1 (M+H)⁺¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.82-8.70 (m, 2H), 8.43 (d, J=4.0Hz, 1H), 8.08 (d, J=9.1 Hz, 1H), 7.93 (d, J=8.2 Hz, 1H), 7.54 (d, J=1.9Hz, 1H), 7.39 (dd, J=10.1, 6.6 Hz, 2H), 6.87 (dd, J=8.6, 5.7 Hz, 2H),5.19 (s, 2H), 3.54 (s, 4H), 2.76 (s, 4H).

Example 73—Synthesis of Compound 073

Step 1:

To a solution of tert-butyl 2-amino-4-(pyridin-2-yl)phenylcarbamate(148.5 mg, 0.52 mmol) and6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic acid (160.7 mg, 0.52mmol) in Py (2.5 ml) was added EDCI (299 mg, 1.56 mmol). The mixture wasstirred at room temperature overnight. The aqueous layer was extractedwith EA (2×100 ml). The combined organic layers were washed with brine(50 ml), dried over Na₂SO₄ and concentrated in vacuum to afford a whitesolid. (240 mg, 100%)

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-2-yl) phenylcarbamoyl)pyridin-2-yl) piperazine-1-carboxylate (240 mg, 0.42 mmol) in DCM (5 ml)at 0° C. was added TFA (2 ml) drop wise. And the resulting mixture wasstirred at room temperature for 2 h. The residue was purified byPrep-HPLC to afford compound 073 as a white solid (97 mg, 62%). LCMS:m/z=375.2 (M+H)⁺ ¹H NMR (400 MHz, DMSO) δ 9.54 (s, 1H), 8.76 (d, J=2.4Hz, 1H), 8.54 (d, J=4.6 Hz, 1H), 8.09 (dd, J=9.0, 2.4 Hz, 1H), 7.95 (d,J=2.0 Hz, 1H), 7.85-7.63 (m, 3H), 7.28-7.10 (m, 1H), 6.92-6.75 (m, 2H),5.26 (s, 2H), 3.65-3.46 (m, 4H), 2.75 (dd, J=27.3, 22.3 Hz, 4H).

Example 74—Synthesis of Compound 074

Step 1:

A mixture of 6-(4-methylpiperazin-1-yl)nicotinic acid (100 mg, 0.45mmol), tert-butyl 2-amino-4-(furan-2-yl)phenylcarbamate (124 mg, 0.45mmol) and EDCI (191 mg, 1 mmol) in pyridine (5 mL) was stirred at roomtemperature for overnight. The mixture was poured into water (20 mL) andextracted with EA to afford crude (270 mg, crude).

Step 2:

A mixture of compound tert-butyl4-(furan-2-yl)-2-(6-(4-methylpiperazin-1-yl)nicotinamido)phenylcarbamate(270 mg, crude) and TFA (3 mL) in DCM (3 mL) was stirred at roomtemperature for 1 h. The mixture was purified by Prep-HPLC to obtaincompound 074 (92 mg, 45%, lot SP-0018110-104) as a yellow solid. LCMS:m/z=378.5 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.51 (s, 1H), 8.76 (s, 1H),8.09 (d, J=7.3 Hz, 1H), 7.58 (d, J=21.9 Hz, 1H), 7.51 (s, 1H), 7.32 (d,J=8.0 Hz, 1H), 6.84 (dt, J=30.4, 15.5 Hz, 2H), 6.55 (dd, J=49.8, 11.4Hz, 2H), 5.15 (s, 2H), 3.63 (s, 4H), 2.44-2.34 (m, 4H), 2.22 (s, 3H).

Example 75—Synthesis of Compound 075

Step 1:

A mixture of 6-(4-methylpiperazin-1-yl)nicotinic acid (50 mg, 0.226mmol), tert-butyl 2-amino-4-(furan-3-yl)phenylcarbamate (62 mg, 0.226mmol) and EDCI (95 mg, 0.5 mmol) in pyridine (5 mL) was stirred at roomtemperature for overnight. The mixture was poured into water (20 mL) andextracted with EA to afford crude (170 mg, crude).

Step 2:

A mixture of compound tert-butyl4-(furan-3-yl)-2-(6-(4-methylpiperazin-1-yl)nicotinamido)phenylcarbamate(170 mg, crude) and TFA (3 mL) in DCM (3 mL) was stirred at roomtemperature for 1 h. The mixture was purified by Prep-HPLC to obtaincompound 075 (63 mg, 61.7%, lot SP-0018110-108) as a yellow solid. LCMS:m/z=378.5 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.53 (s, 1H), 8.75 (s, 1H),8.10 (d, J=8.8 Hz, 1H), 7.96 (s, 1H), 7.66 (s, 1H), 7.36 (s, 1H), 7.23(d, J=8.3 Hz, 1H), 6.97-6.71 (m, 3H), 4.97 (s, 2H), 3.62 (s, 4H),2.43-2.35 (m, 4H), 2.22 (s, 3H).

Example 76—Synthesis of Compound 076

Step 1:

A mixture of 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic acid(184 mg, 0.6 mmol), tert-butyl 3-amino-2′-fluorobiphenyl-4-ylcarbamate(180 mg, 0.54 mmol) and EDCI (229 mg, 1.2 mmol) in pyridine (10 mL) wasstirred at room temperature for overnight. The mixture was poured intowater (20 mL) and extracted with EA, then purified by preparative TLC(PE/EA=2:1) to afford desired compound (210 mg, 66%) as a pale solid.

Step 2:

A mixture of compound tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-2′-fluorobiphenyl-3-ylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate(200 mg, 0.33 mmol) and TFA (2 mL) in DCM (5 mL) was stirred at roomtemperature for 1 h. The mixture was concentrated, ether was added,basified with saturated NaHCO₃, the solid was collected to obtaincompound 076 (54 mg, 88%, lot SP-0018110-116) as a yellow solid. LCMS:m/z=392.4 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.52 (s, 1H), 8.75 (d, J=1.8Hz, 1H), 8.08 (d, J=7.1 Hz, 1H), 7.54-7.12 (m, 7H), 6.96-6.71 (m, 2H),5.16 (s, 2H), 3.57 (dd, J=20.8, 16.4 Hz, 4H), 2.87-2.70 (m, 4H).

Example 77—Synthesis of Compound 077

Step 1:

The title compound tert-butyl 3′-fluoro-3-nitrobiphenyl-4-ylcarbamatewas synthesized following the same procedures as described in Example 3.

Step 2:

A mixture of 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic acid(100 mg, 0.3 mmol), tert-butyl 3′-fluoro-3-nitrobiphenyl-4-ylcarbamate(90 mg, 0.27 mmol) and EDCI (115 mg, 0.6 mmol) in pyridine (8 mL) wasstirred at room temperature for overnight. The mixture was poured intowater (20 mL) and extracted with EA, then purified by preparative TLC(PE/EA=2:1) to afford desired compound (98 mg, 50%) as a pale solid.

Step 3:

A mixture of compound tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-3′-fluorobiphenyl-3-ylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate(98 mg, 0.166 mmol) and TFA (2 mL) in DCM (5 mL) was stirred at roomtemperature for 1 h. The mixture was concentrated, ether was added,basified with saturated NaHCO₃, the solid was collected to obtaincompound 077 (110 mg, 83%, lot SP-0017467-135) as a pale yellow solid.LCMS: m/z=392.4 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.57 (s, 1H), 8.75 (s,1H), 7.55 (s, 1H), 7.39 (dd, J=24.0, 7.5 Hz, 5H), 7.05 (s, 1H),6.93-6.80 (m, 3H), 5.20 (s, 3H), 3.55 (s, 4H), 2.77 (s, 4H).

Example 78—Synthesis of Compound 078

Step 1:

The title compound tert-butyl3-amino-3′,5′-difluorobiphenyl-4-ylcarbamate was synthesized followingthe same procedures as described in Example 3.

Step 2:

A mixture of 6-(4-(tert-butoxycarbonyl)piperazin-1-yl)nicotinic acid(307 mg, 1 mmol), tert-butyl3-amino-3′,5′-difluorobiphenyl-4-ylcarbamate (288 mg, 0.9 mmol) and EDCI(382 mg, 2 mmol) in pyridine (10 mL) was stirred at room temperature forovernight. The mixture was poured into water (20 mL) and extracted withEA, then purified by preparative TLC (PE/EA=2:1) to afford desiredcompound (360 mg, 59%) as a pale solid.

Step 3:

A mixture of compound tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-3′,5′-difluorobiphenyl-3-ylcarbamoyl)pyridin-2-yl)piperazine-1-carboxylate(360 mg, 0.59 mmol) and TFA (3 mL) in DCM (10 mL) was stirred at roomtemperature for 1 h. The mixture was concentrated, basified withsaturated NaHCO₃, then extracted with EA, concentrated to afford thecrude, washed with 5 ml EA to obtain compound 078 (50 mg, 21%, lotSP-0018270-024) as a yellow solid. LCMS: m/z=410.5 (M+H)⁺. ¹H NMR (400MHz, DMSO) δ 9.51 (s, 1H), 8.76 (s, 1H), 8.09 (d, J=7.1 Hz, 1H),7.72-6.67 (m, 7H), 5.28 (s, 2H), 3.55 (s, 4H), 2.77 (s, 4H).

Example 79—Synthesis of Compound 079

Step 1:

The title compound 5-(4-methylpiperazin-1-yl)picolinic acid (lithiumsalt) was synthesized following the same procedures as described inExample 28.

Step 2:

A mixture of lithium 5-(4-methylpiperazin-1-yl)picolinate (0.60 mmol)and HATU (252 mg, 0.66 mmol) in DMF (6 mL) was stirred at roomtemperature for 20 min. Then tert-butyl 3-aminobiphenyl-4-ylcarbamate(180 mg, 0.63 mmol) was added. It was stirred at room temperature for 2h. Then it was poured into water. The resulting solid was collected byfiltration and dried in vacuo to afford desired compound.

Step 3: To a mixture of tert-butyl 3-(5-(4-methylpiperazin-1-yl)picolinamido) biphenyl-4-ylcarbamate (0.60 mmol) in MeOH (2 mL) wasadded HCl/dioxane (4 mL) at 0° C. It was stirred at 0° C. to ambienttemperature for 18 h. It was concentrated in vacuo and the residue waspurified by Prep-HPLC to give product as a white solid as compound 079(117 mg, yield: 50%, three steps, lot: SP-0017146-021). LCMS: m/z=388.2(M+H)⁺. ¹H NMR (500 MHz, DMSO) δ 9.89 (s, 1H), 8.40 (d, J=2.8 Hz, 1H),7.96 (d, J=8.8 Hz, 1H), 7.89 (d, J=2.1 Hz, 1H), 7.56 (d, J=7.3 Hz, 2H),7.48 (dd, J=8.9, 2.9 Hz, 1H), 7.40 (t, J=7.7 Hz, 2H), 7.30-7.21 (m, 2H),6.91 (d, J=8.3 Hz, 1H), 5.04 (s, 2H), 3.40-3.36 (m, 4H), 2.49-2.45 (m,4H), 2.24 (s, 3H).

Example 80—Synthesis of Compound 080

Step 1:

A mixture of lithium 5-(4-methylpiperazin-1-yl)picolinate (0.60 mmol)and HATU (252 mg, 0.66 mmol) in DMF (4 mL) was stirred at roomtemperature for 15 min. Then tert-butyl2-amino-4-(thiophen-2-yl)phenylcarbamate (192 mg, 0.66 mmol) was added.It was stirred at room temperature for 2 h. Then it was poured intowater. The resulting solid was collected by filtration and dried invacuo to give crude compound.

Step 2:

To a mixture of tert-butyl 3-(5-(4-methylpiperazin-1-yl) picolinamido)biphenyl-4-ylcarbamate (0.60 mmol) in MeOH (4 mL) was added HCl/dioxane(4 mL) at 0° C. It was stirred at 0° C. to ambient temperature for 18 h.It was concentrated in vacuo and the residue was purified by prep-HPLCto give compound 080 as a white solid (87 mg, yield: 29%, three steps,lot SP-0017146-020). LCMS: m/z=394.2 (M+H)⁺. ¹H NMR (500 MHz, DMSO) δ9.87 (s, 1H), 8.39 (d, J=2.8 Hz, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.87 (d,J=2.1 Hz, 1H), 7.47 (dd, J=8.9, 2.8 Hz, 1H), 7.37 (d, J=4.4 Hz, 1H),7.31-7.19 (m, 2H), 7.06 (dd, J=5.0, 3.6 Hz, 1H), 6.85 (d, J=8.3 Hz, 1H),5.08 (s, 2H), 3.40-3.35 (m, 4H), 2.49-2.45 (m, 4H), 2.24 (s, 3H).

Example 81—Synthesis of Compound 081

Step 1:

To a solution of tert-butyl 4-bromo-2-nitrophenylcarbamate (8.0 g, 25.2mmol) and 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane)(7.7 g, 30.3 mmol), KOAc 5.0 g, 51.0 mmol), PdCl₂ (dppf-DCM) (922 mg,1.3 mmol) in 1,4-dioxane (50 ml) under N₂ protection. The mixture wasstirred at 80° C. overnight. The mixture was directly purified on columnwith (PE/EA=50/1) to afford a yellow solid (7.8 g, 80%).

Step 2:

To a solution of tert-butyl2-nitro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylcarbamate(3.0 g, 8.24 mmol) and 2-bromopyridine (1.3 g, 8.24 mmol) in 1,4-dioxane(15 ml) and H₂O (8 ml) was added K₂CO₃ (1.7 g, 16.48 mmol) and Pd(PPh3)₄(476 mg, 0.41 mmol). The mixture was stirred at 100° C. for overnight.The mixture was directly purified on column with (PE/EA=20/1) to afforda yellow solid (1 g, 38%).

Step 3:

To a solution of tert-butyl 2-nitro-4-(pyridin-2-yl)phenylcarbamate (1.5g, 4.76 mmol) and FeCl₃ (155 mg, 0.95 mmol) in EtOH (15 ml) was addedCarbon (1.5 g, 125 mmol) and drop wise N₂H₄H2O (8 ml). The mixture wasstirred at 60° C. for 2 h. The aqueous layer was extracted with EA (100ml). The combined organic layers were washed with brine (50 ml), driedover Na₂SO₄, filtered and concentrated in vacuum to afford a gray solid.(1.1 g, 76%)

Step 4:

To a solution of 5-(4-(tert-butoxycarbonyl)piperazin-1-yl)picolinic acid(216 mg, 0.62 mmol) and HATU (232 mg, 0.62 mmol) in DMF (7 ml) that wasstirred at room temperature for 15 min., tert-butyl2-amino-4-(pyridin-2-yl)phenylcarbamate (100 mg, 0.35 mmol) was added.The mixture was stirred at room temperature for overnight. Water wasadded at 0° C. The resulting precipitate was collected by filtration anddried in vacuum to afford a white solid (200 mg, 100%).

Step 5:

To a solution of tert-butyl4-(6-(2-(tert-butoxycarbonylamino)-5-(pyridin-2-yl) phenylcarbamoyl)pyridin-3-yl) piperazine-1-carboxylate (200 mg, 0.35 mmol) in DCM (2 ml)at 0° C. was added TFA (2 ml) dropwise. The resulting solution wasstirred at room temperature for 2 h. The crude product was purified byPrep-HPLC to afford a white solid as compound 081 (53 mg, 41% yield, lotSP-0018108-028). LCMS: m/z=375.2 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.83(s, 1H), 8.55 (d, J=4.7 Hz, 1H), 8.37 (d, J=2.8 Hz, 1H), 8.27 (d, J=2.0Hz, 1H), 7.95 (d, J=8.8 Hz, 1H), 7.77 (d, J=3.4 Hz, 2H), 7.72-7.65 (m,1H), 7.45 (d, J=6.0 Hz, 1H), 7.20 (dd, J=8.6, 4.6 Hz, 1H), 6.88 (d,J=8.4 Hz, 1H), 5.21 (s, 2H), 3.30-3.26 (m, 4H), 2.87-2.81 (m, 4H).

Example 82—Synthesis of Compound 082

Step 1:

A mixture of lithium 5-(4-methylpiperazin-1-yl)picolinate (176 mg, 0.63mmol) and HATU (240 mg, 0.63 mmol) in DMF (5 mL) was stirred at roomtemperature for 15 min. Then tert-butyl 2-amino-4-(furan-2-yl)phenylcarbamate (96 mg, 0.35 mmol) was added. It was stirred at roomtemperature for 18 h. Water was added. The resulting solid was collectedby filtration and dried in vacuo to give crude compound.

Step 2:

To a mixture of above tert-butyl4-(furan-2-yl)-2-(5-(4-methylpiperazin-1-yl) picolinamido)phenylcarbamate (0.35 mmol) in DCM (2 mL) was added TFA (2 mL) at 0° C.It was stirred at 0° C. and then at room temperature until the reactionwas complete. It was concentrated in vacuo and the residue was purifiedby Prep-HPLC to give compound 082 as a white solid (57.0 mg, free amine,yield: 43%, two steps, lot SP-0017146-145). LCMS: m/z=378.1 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.84 (s, 1H), 8.39 (d, J=2.6 Hz, 1H), 7.93 (dd,J=17.9, 5.2 Hz, 2H), 7.62 (s, 1H), 7.47 (dd, J=8.8, 2.7 Hz, 1H), 7.29(dd, J=8.3, 1.9 Hz, 1H), 6.85 (d, J=8.3 Hz, 1H), 6.61 (d, J=3.2 Hz, 1H),6.51 (dd, J=3.2, 1.8 Hz, 1H), 5.08 (s, 2H), 3.42-3.35 (m, 4H), 2.49-2.42(m, 4H), 2.24 (s, 3H).

Example 83—Synthesis of Compound 083

Step 1:

To a solution of 5-(4-methylpiperazin-1-yl) picolinic acid (170 mg, 0.77mmol) and HATU (292 mg, 0.77 mmol) in DMF (5 ml) that was stirred atroom temperature for 15 min., tert-butyl2-amino-4-(furan-3-yl)phenylcarbamate (117 mg, 0.43 mmol) was added. Themixture was stirred at room temperature for overnight. The mixture wasadded to water, then a white solid formed. It was filtered through aCelite pad, and the filtrate was concentrated to afford a yellow solid(154 mg, 92.2% yield).

Step 2:

To a solution of tert-butyl 4-(furan-3-yl)-2-(5-(4-methylpiperazin-1-yl)picolinamido) phenylcarbamate (154 mg, 0.32 mmol) in DCM (2 ml) at 0° C.was added TFA (2 ml) dropwise. And the resulted was stirred at roomtemperature for 2 h. The residue mixture was concentrated in vacuum. Theresidue was purified by Prep-HPLC. The white solid was re-crystallizedfrom water, dried by lyophilization to afford white solid as compound083. (62 mg, 51.7% yield, lot SP-0018108-083). LCMS: m/z=378.2 (M+H)⁺.¹H NMR (400 MHz, DMSO) δ 9.83 (s, 1H), 8.39 (d, J=2.7 Hz, 1H), 8.00-7.89(m, 2H), 7.74 (d, J=1.7 Hz, 1H), 7.67 (s, 1H), 7.48 (dd, J=8.9, 2.8 Hz,1H), 7.20 (dd, J=8.2, 1.8 Hz, 1H), 6.88-6.76 (m, 2H), 4.93 (s, 2H),3.42-3.35 (m, 4H), 2.49-2.45 (m, 4H).

Example 84—Synthesis of Compound 084

Step 1:

A mixture of 5-(4-(tert-butoxycarbonyl) piperazin-1-yl) picolinic acid(100 mg, 0.33 mmol), tert-butyl3-amino-3′,5′-difluorobiphenyl-4-ylcarbamate (106 mg, 0.33 mmol) andEDCI (190 mg, 0.99 mmol) in Py (3 ml) was stirred at room temperaturefor overnight. It was extracted with EA and purified by columnchromatography (PE:EA=2:1). White solid was afforded (175 mg, 69%yield).

Step 2:

To a solution of tert-butyl4-(6-(4-(tert-butoxycarbonylamino)-3′,5′-difluorobiphenyl-3-ylcarbamoyl)pyridin-3-yl) piperazine-1-carboxylate (175 mg, 0.29 mmol) in DCM (4 ml)was added TFA (2 ml). The mixture was stirred at room temperature for 2h. It was concentrated and neutralized by aqueous NaOH (2 mol/L). Whitesolid was filtered as compound 084. (85 mg, 72% yield, lotSP-0018421-087). LCMS: m/z=410.2 (M+H)⁺. ¹H NMR (400 MHz, DMSO) δ 9.87(s, 1H), 8.38 (d, J=2.6 Hz, 1H), 7.96 (d, J=8.8 Hz, 1H), 7.89 (d, J=2.0Hz, 1H), 7.46 (dd, J=8.7, 2.7 Hz, 1H), 7.38 (dd, J=8.4, 2.2 Hz, 1H),7.29 (d, J=7.4 Hz, 2H), 7.08 (t, J=9.2 Hz, 1H), 6.89 (d, J=8.4 Hz, 1H),5.22 (s, 2H), 2.94-2.84 (m, 4H), 2.58 (m, 4H).

Example 85—Synthesis of Compound 085

Step 1:

A mixture of methyl 5-chloropyrazine-2-carboxylate (1.72 g, 10 mmol),tert-butyl piperazine-1-carboxylate (3.72 g, 20 mmol) and DIPEA (3.87 g,30 mmol) in 1,4-dioxane (20 mL) was stirred at 100° C. for overnight.The mixture was concentrated to get a residue, which was purified bysilica gel to get compound (3.06 g, 95%).

Step 2:

A mixture of methyl methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazine-2-carboxylate (322 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 3-aminobiphenyl-4-ylcarbamate (284 mg, 1 mmol), EDCI (573 mg,3 mmol) and Py (10 mL). The mixture was stirred at room temperature forovernight. The mixture was poured into water (50 mL) and filtered toafford compound (459 mg, 80%).

Step 3:

To a solution of tert-butyl4-(5-(4-(tert-butoxycarbonylamino)biphenyl-3-ylcarbamoyl)pyrazin-2-yl)piperazine-1-carboxylate(287 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to get compound 085 (131 mg, 70%). LCMS: m/z=375.2 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.70 (s, 1H), 8.72 (s, 1H), 8.34 (s, 1H), 7.81 (s,1H), 7.56 (d, J=7.7 Hz, 2H), 7.40 (t, J=7.6 Hz, 2H), 7.32-7.20 (m, 2H),6.90 (d, J=8.3 Hz, 1H), 5.06 (s, 2H), 3.72-3.59 (m, 4H), 2.85-2.74 (m,4H).

Example 86—Synthesis of Compound 086

Step 1:

A mixture of methyl 5-chloropyrazine-2-carboxylate (1.72 g, 10 mmol),tert-butyl piperazine-1-carboxylate (2.0 g, 20 mmol) and DIPEA (3.87 g,30 mmol) in 1,4-dioxane (20 mL) was stirred at 100° C. for overnight.The mixture was concentrated to get a residue, which was purified bysilica gel to get compound (1.65 g, 70%).

Step 2:

A mixture of methyl 5-(4-methylpiperazin-1-yl)pyrazine-2-carboxylate(236 mg, 1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirredat 60° C. for 3 h. The mixture was concentrated to get a residue, whichwas added tert-butyl 2-amino-4-(thiophen-2-yl)phenylcarbamate (290 mg, 1mmol), EDCI (573 mg, 3 mmol) and Py (10 mL). The mixture was stirred atroom temperature for overnight. The mixture was poured into water (50mL), filtered to get compound (420 mg, 85%).

Step 3:

To a solution of tert-butyl2-(5-(4-methylpiperazin-1-yl)pyrazine-2-carboxamido)-4-(thiophen-2-yl)phenylcarbamate(247 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to get compound 086 (138 mg, 70%). LCMS: m/z=395.1 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.69 (s, 1H), 8.73 (s, 1H), 8.37 (s, 1H), 7.78 (d,J=1.7 Hz, 1H), 7.37 (d, J=5.0 Hz, 1H), 7.26 (dd, J=11.4, 2.8 Hz, 2H),7.09-6.97 (m, 1H), 6.84 (d, J=8.3 Hz, 1H), 5.10 (s, 2H), 3.80-3.66 (m,4H), 2.46-2.37 (m, 4H), 2.23 (s, 3H).

Example 87—Synthesis of Compound 087

Step 1:

A mixture of methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazine-2-carboxylate (236 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 2-amino-4-(furan-2-yl)phenylcarbamate (274 mg, 1 mmol), EDCI(573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to get compound (451 mg, 80%)

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-2-yl)phenylcarbamoyl)pyrazin-2-yl)piperazine-1-carboxylate(282 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to get compound 087 (146 mg, 80%) LCMS: m/z=365.2 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.65 (s, 1H), 8.71 (s, 1H), 8.34 (s, 1H), 7.83 (s,1H), 7.62 (s, 1H), 7.30 (d, J=7.4 Hz, 1H), 6.84 (d, J=8.3 Hz, 1H), 6.61(d, J=2.7 Hz, 1H), 6.51 (s, 1H), 5.10 (s, 2H), 3.66 (s, 4H), 2.80 (s,4H).

Example 88—Synthesis of Compound 088

Step 1:

A mixture of methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazine-2-carboxylate (236 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 2-amino-4-(furan-3-yl)phenylcarbamate (274 mg, 1 mmol), EDCI(573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to get compound 3 (451 mg, 80%)

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(furan-3-yl)phenylcarbamoyl)pyrazin-2-yl)piperazine-1-carboxylate(282 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to get compound 088 (146 mg, 80%). LCMS: m/z=365.1 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.66 (s, 1H), 8.71 (s, 1H), 8.33 (s, 1H), 7.96 (s,1H), 7.67 (s, 2H), 7.21 (d, J=8.2 Hz, 1H), 6.87-6.77 (m, 2H), 4.95 (s,2H), 3.66 (s, 4H), 2.80 (s, 4H).

Example 89—Synthesis of Compound 089

Step 1:

A mixture of methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazine-2-carboxylate (236 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 2-amino-4-(pyridin-4-yl)phenylcarbamate (285 mg, 1 mmol),EDCI (573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to get compound (401 mg, 70%)

Step 2:

To a solution of tert-butyl4-(5-(2-(tert-butoxycarbonylamino)-5-(pyridin-4-yl)phenylcarbamoyl)pyrazin-2-yl)piperazine-1-carboxylate(288 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to get compound 089 (150 mg, 80%). LCMS: m/z=376.1 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.70 (s, 1H), 8.72 (s, 1H), 8.52 (d, J=5.6 Hz,2H), 8.34 (s, 1H), 7.91 (s, 1H), 7.58 (d, J=5.7 Hz, 2H), 7.48 (d, J=8.5Hz, 1H), 6.91 (d, J=8.4 Hz, 1H), 5.32 (s, 2H), 3.66 (s, 4H), 2.80 (s,4H).

Example 90—Synthesis of Compound 090

Step 1:

A mixture of methyl5-(4-(tert-butoxycarbonyl)piperazin-1-yl)pyrazine-2-carboxylate (236 mg,1 mmol) and LiOH (126 mg, 3 mmol) in MeOH (20 mL) was stirred at 60° C.for 3 h. The mixture was concentrated to get a residue, which was addedtert-butyl 3-amino-4′-fluorobiphenyl-4-ylcarbamate (302 mg, 1 mmol),EDCI (573 mg, 3 mmol) and Py (10 mL). The mixture was stirred at roomtemperature for overnight. The mixture was poured into water (50 mL),filtered to get compound (444 mg, 75%).

Step 2

To a solution of tert-butyl4-(5-(4-(tert-butoxycarbonylamino)-4′-fluorobiphenyl-3-ylcarbamoyl)pyrazin-2-yl)piperazine-1-carboxylate(296 mg, 0.5 mmol) in DCM (5 mL) was added TFA (2.5 mL), stirred for 1h. The mixture was concentrated to get a residue, which was purified byPrep-HPLC to get compound 090 (157 mg, 80%). LCMS: m/z=393.2 (M+H)⁺. ¹HNMR (400 MHz, DMSO) δ 9.94 (s, 1H), 9.04 (s, 2H), 8.79 (d, J=1.1 Hz,1H), 8.46 (s, 1H), 7.79 (d, J=2.1 Hz, 1H), 7.61 (dd, J=8.8, 5.5 Hz, 2H),7.35 (dd, J=8.3, 2.1 Hz, 1H), 7.25 (t, J=8.9 Hz, 2H), 7.03 (d, J=8.3 Hz,1H), 4.02-3.92 (m, 4H), 3.26 (s, 4H).

Example 91—Pharmacokinetics

Male SD rats were fasted overnight. Compounds of the invention weredissolved in dimethyl acetamide at 10 times the final concentration,then Solutol HS 15 (BASF) was added to a final concentration of 10%.Finally 80% saline was added and vortexed to achieve a clear solution.For the IV dosing three animals were injected via the foot dorsal veinwith 1 mg/kg compound. For the PO dosing 5 mg/kg of compound wasdelivered by oral gavage. Blood was collected via the tail vein intoK2EDTA tubes at 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 4hours, 8 hours and 24 hours after dosing. The blood was centrifuged at2000 g for 5 minutes at 4° C. to obtain plasma. The plasma was extractedwith acetonitrile and the level of compound was analyzed by LC/MS/MS.The level of compound in plasma was calculated from a standard curve inrat plasma. The IV clearance (L/h/kg) and area under the curve (h*ng/mL)were calculated using WinNonLin software. The dose adjusted area underthe curve for the IV and oral dosing were used to calculate the oralbioavailability.

Pharmacokinetic properties were assessed in a rat cassette dosingexperiment. The IV clearance (IV Clr.) is in units of L/hr/kg. The oralmaximum plasma concentration (PO Cmax) is in units of ng/ml. The oralplasma half-life (PO T½) is in units of hours. The oral area under thecurve (PO AUC) is in units of hours*ng/ml. The fraction absorbed by theoral route (F %) is a percentage of the oral area under the curve to theIV area under the curve, dose adjusted. A summary of results ispresented in Table 2, below.

TABLE 2 Compound IV PO PO PO No. Structure Clr. T½ Cmax AUC F% 002

0.12 5.9 3746 34597 86.1 003

0.19 5.0 1910 19108 77.0 004

0.04 4.3 10353 91943 90.8 008

0.86 5.6 536 5536 100 010

1.1 4.7 164 570 12.6 012

0.53 7.7 534 8186 93.9 013

0.78 8.1 425 5783 105 014

0.66 4.9 727 7097 97.0 021

5.66 3.2 68 763 89.8 022

0.98 13.5 517 8183 195 023

0.36 8.5 664 6368 51.2 024

0.21 4.4 1000 8225 36.3 027

0.63 3.0 839 6191 78.6 030

0.48 12.8 944 15260 170 032

1.06 4.3 173 456 9.7 033

0.65 13.7 614 10714 191 035

0.17 4.4 1870 20150 70.5 036

0.42 7.3 500 7282 69.1 040

1.6 16.7 211 3338 151

Example 92—hERG and Cyp Inhibition

For hERG inhibition assays, the cells used were HEK293 cells stablytransfected with hERG (cell line obtained from Cytocentrics Inc. 3463Magic Drive San Antonio, Tex. 78229). Composition of External Solution:NaCl, 137 mM; KCl, 4 mM; MgCl2, 1.0 mM; CaCl₂), 1.8 mM; HEPES 10 mM;Dextrose 11 mM; Adjusted to a pH of 7.4 with NaOH. Composition ofInternal Solution: KCl, 130.0 mM; MgCl2, 1.0 mM; HEPES, 5.0 mM; EGTA,5.0 mM; NaCl 7.0 mM. Adjusted to a pH of 7.2 using KOH. TestConcentrations: 0.1, 1, 10, 100 μM. Vehicle: DMSO. Experiments wereperformed at 34+/−1° C. Current was recorded using the whole-cell patchclamp technique on the Cytopatch automated platform. hERG current waselicited with the following voltage protocol: hERG current amplitude wasmeasured as the peak current at −50 mV (tail current). The percentblockade of hERG was measured as current reduction after a steady-stateeffect had been reached in the presence of drug relative to currentamplitude before drug was introduced (control). Each cell served as itsown control. Data are presented as the 1050 calculated by non-linearregression analysis.

For Cyp inhibition, human liver microsomes from BD Gentest wereincubated with Compound 028 or Compound 032 (10, 3.33, 1.11, 0.37, 0.12,0.04, 0.01 μM) and substrate (CYP1A2: Phenacetin at 30 μM; CYP2C9:Diclofenac at 10 μM; CYP2C19: S-Mephenytoin at 35 μM; CYP3A4: Midazolamat 5 μM and Testosterone at 80 μM; CYP2D6: Bufuralol at 10 μM) for thefollowing incubation times: CYP1A2, 2C9, 2D6: 10 minutes, 37° C.;CYP2C19: 45 minutes, 37° C.; CYP3A4: 5 minutes, 37° C. Substrateconversion was measured by liquid chromatography/mass spectrometry/massspectrometry (LC/MS/MS). Inhibition was calculated by curve fitting inGraph Pad Prism.

A summary of results is presented in Table 3, below. A—indicates thatthe compound was not tested.

TABLE 3 Compound hERG Cyp ID Structure (IC₅₀) (IC₅₀) 001

>30 μM — 003

7.6 μM >10 μM (all isozymes) 006

5.1 μM >10 μM (all isozymes) 007

8.6 μM >10 μM (all isozymes) 021

4.2 μM — 022

0.4 μM >10 μM (all isozymes) 025

8.9 μM >10 μM (all isozymes) 026

>100 μM — 027

>100 μM >10 μM (all isozymes) 028

38 μM 5.9 μM (3A4/midazolam) 029

5.2 μM — 030

70 μM >10 μM (all isozymes) 031

12 μM >10 μM (all isozymes) 032

12 μM 9.5 μM (3A4/midazolam) 038

2.5 — 039

3.6 —

Example 93—HDAC Enzyme Assays

Compounds for testing were diluted in DMSO to 50 fold the finalconcentration and a ten-point three-fold dilution series was made. Thecompounds were diluted in assay buffer (50 mM HEPES, pH 7.4, 100 mM KCl,0.001% Tween-20, 0.05% BSA, 20 μM TCEP) to 6-fold their finalconcentration. The HDAC enzymes (purchased from BPS Biosciences) werediluted to 1.5-fold their final concentration in assay buffer. Thetripeptide substrate and trypsin at 0.05 μM final concentration werediluted in assay buffer at 6-fold their final concentration. The finalenzyme concentrations used in these assays were 3.3 ng/ml (HDAC1), 0.2ng/ml (HDAC2) and 0.08 ng/ml (HDAC3). The final substrate concentrationsused were 16 μM (HDAC1), 10 μM (HDAC2) and 17 μM (HDAC3).

Five μl of compounds and 20 μl of enzyme were added to wells of a black,opaque 384 well plate in duplicate. Enzyme and compound were incubatedtogether at room temperature for 10 minutes. Five μl of substrate wasadded to each well, the plate was shaken for 60 seconds and placed intoa Victor 2 microtiter plate reader. The development of fluorescence wasmonitored for 60 min and the linear rate of the reaction was calculated.The IC₅₀ was determined using Graph Pad Prism by a four parameter curvefit. In Table 4 below, letter designations are defined as follows:A=IC₅₀ value <10 nM, B=IC₅₀ value between 10 and 500 nM, C=IC₅₀value >500 nM.

TABLE 4 Compound ID Structure HDAC1 HDAC2 HDAC3 034

A B B 035

A B B 036

A B B 037

A B B 038

A B B 039

A A B 040

A A B 041

A B B 042

A B B 043

A B C 044

B B C 045

B B B 046

A B B 047

A B C 048

A B C 049

A B B 050

A B B 051

A A B 052

A B B 053

A A B 054

A B B 055

C B C 056

B B C 057

C B C 058

B B B 059

C B B 060

C C C 061

C C C 062

C C C 063

C C C 064

A B B 065

A B B 066

A B B 067

A B B 068

A B B 069

B B C 070

A B B 071

A B B 072

B B B 073

B B C 074

A B B 075

A B B 076

A B B 077

B B B 078

C B C 079

A B B 080

A A B 081

B B C 082

A B B 083

A B B 084

C C C 085

A B B 086

A A B 087

A B B 088

A B B 089

B B B 090

A B B

1-21. (canceled)
 22. A method for treating sickle cell disease, betathalassemia, myelodysplastic syndrome, acute myelogenous leukemia,neuroblastoma, or acute megakaryocytic leukemia in a subject in needthereof, comprising administering to the subject a therapeuticallyeffective amount of a compound of Formula III:

or a pharmaceutically acceptable salt thereof; wherein (a) X is C(H), Yis C(H), and Z is N; R⁵ is H or phenyl; and R⁶ is H; or (b) X is C(H), Yis C(H), and Z is N; R⁵ is 1-furanyl; and R⁶ is C₁₋₄ alkyl, wherein theC₁₋₄ alkyl group is optionally, independently substituted one or moretimes with halo, —CN, —NO₂, —C₁-C₆ alkoxy, —C₁-C₆ alkyl, —C₁-C₆haloalkyl, and C(O)—C₁-C₆ alkyl; or (c) X is C(H), Y is N, and Z isC(H); R⁵ is H, 1-furanyl, 2-furanyl, pyridinyl, or thiophenyl; and R⁶ isH; or (d) X is N, Y is C(H), and Z is C(H); R⁵ is H, 1-furanyl,2-furanyl, pyridinyl, or phenyl, wherein phenyl is optionallysubstituted with halogen; and R⁶ is H.
 23. (canceled)
 24. The method ofclaim 22, wherein X is C(H), Y is C(H), and Z is N; R⁵ is 1-furanyl; andR⁶ is CH₃.
 24. The method of claim 22, wherein X is C(H), Y is C(H), andZ is N; R⁵ is H; and R⁶ is H.
 25. The method of claim 22, wherein thecompound of Formula III is:

or a pharmaceutically acceptable salt thereof.
 26. The method of claim12, wherein the method treats sickle cell disease.
 27. A method fortreating sickle cell disease, beta thalassemia, myelodysplasticsyndrome, acute myelogenous leukemia, neuroblastoma, or acutemegakaryocytic leukemia in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of acompound of compound of Formula IV:

or a pharmaceutically acceptable salt thereof; wherein R⁷ is furanyl,pyridinyl, or thiophenyl.
 28. The method of claim 27, wherein thecompound of Formula IV is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 29. The method of claim27, wherein the method treats sickle cell disease.
 30. A method fortreating lung cancer, colon cancer, breast cancer, neuroblastoma,leukemia, lymphoma, or neuroblastoma in a subject in need thereof,comprising administering to the subject a therapeutically effectiveamount of a compound of compound of Formula III:

or a pharmaceutically acceptable salt thereof; wherein (a) X is C(H), Yis C(H), and Z is N; R⁵ is H or phenyl; and R⁶ is H; or (b) X is C(H), Yis C(H), and Z is N; R⁵ is 1-furanyl; and R⁶ is 01-4 alkyl, wherein theC₁₋₄ alkyl group is optionally, independently substituted one or moretimes with halo, —CN, —NO₂, —C₁-C₆ alkoxy, —C₁-C₆ alkyl, —C₁-C₆haloalkyl, and C(O)—C₁-C₆ alkyl; or (c) X is C(H), Y is N, and Z isC(H); R⁵ is H, 1-furanyl, 2-furanyl, pyridinyl, or thiophenyl; and R⁶ isH; or (d) X is N, Y is C(H), and Z is C(H); R⁵ is H, 1-furanyl,2-furanyl, pyridinyl, or phenyl, wherein phenyl is optionallysubstituted with halogen; and R⁶ is H.
 31. The method of claim 30,wherein X is C(H), Y is C(H), and Z is N; R⁵ is 1-furanyl; and R⁶ isCH₃.
 32. The method of claim 30, wherein X is C(H), Y is C(H), and Z isN; R⁵ is H; and R⁶ is H.
 33. The method of claim 30, wherein thecompound of Formula III is:

or a pharmaceutically acceptable salt thereof.
 34. The method of claim30, wherein the cancer is neuroblastoma.
 35. The method of claim 30,wherein the leukemia is acute myelogenous leukemia or acutemegakaryocytic leukemia.
 36. A method for treating lung cancer, coloncancer, breast cancer, neuroblastoma, leukemia, lymphoma, orneuroblastoma in a subject in need thereof, comprising administering tothe subject a therapeutically effective amount of a compound of compoundof Formula IV:

or a pharmaceutically acceptable salt thereof; wherein R⁷ is furanyl,pyridinyl, or thiophenyl.
 37. The new of claim 36, wherein the compoundof Formula IV is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 38. The method of claim36, wherein the cancer is neuroblastoma.
 39. The method of claim 36,wherein the leukemia is acute myelogenous leukemia or acutemegakaryocytic leukemia.